Spiro Bicyclic Inhibitors Of Menin-MLL Interaction

ABSTRACT

The present invention relates to pharmaceutical agents useful for therapy and/or prophylaxis in a mammal, and in particular to spiro bicyclic compounds, pharmaceutical composition comprising such compounds, and their use as menin/MLL protein/protein interaction inhibitors, useful for treating diseases such as cancer, myelodysplastic syndrome (MDS) and diabetes.

FIELD OF THE INVENTION

The present invention relates to pharmaceutical agents useful fortherapy and/or prophylaxis in a mammal, and in particular to spirobicyclic compounds, pharmaceutical composition comprising suchcompounds, and their use as menin/MLL protein/protein interactioninhibitors, useful for treating diseases such as cancer, myelodysplasticsyndrome (MDS) and diabetes.

BACKGROUND OF THE INVENTION

Chromosomal rearrangements affecting the mixed lineage leukemia gene(MLL; MLL1; KMT2A) result in aggressive acute leukemias across all agegroups and still represent mostly incurable diseases emphasizing theurgent need for novel therapeutic approaches. Acute leukemias harboringthese chromosomal translocations of MLL represent as lymphoid, myeloidor biphenotypic disease and constitute 5 to 10% of acute leukemias inadults and approximately 70% in infants (Marschalek, Br J Haematol 2011.152(2), 141-54; Tomizawa et al., Pediatr Blood Cancer 2007. 49(2),127-32).

MLL is a histone methyltransferase that methylates histone H3 on lysine4 (H3K4) and functions in multiprotein complexes. Use of inducibleloss-of-function alleles of Mll1 demonstrated that Mll1 plays anessential role in sustaining hematopoietic stem cells (HSCs) anddeveloping B cells although its histone methyltransferase activity isdispensable for hematopoiesis (Mishra et al., Cell Rep 2011. 7(4),1239-47).

Fusion of MLL with more than 60 different partners has been reported todate and has been associated with leukemia formation/progression (Meyeret al., Leukemia 2013. 27, 2165-2176). Interestingly, the SET(Su(var)3-9, enhancer of zeste, and trithorax) domain of MLL is notretained in chimeric proteins but is replaced by the fusion partner(Thiel et al., Bioessays 2012. 34, 771-80). Recruitment of chromatinmodifying enzymes like Dot1L and/or the pTEFb complex by the fusionpartner leads to enhanced transcription and transcriptional elongationof MLL target genes including HOXA genes (e.g. HOXA9) and the HOXcofactor MEIS1 as the most prominent ones. Aberrant expression of thesegenes in turn blocks hematopoietic differentiation and enhancesproliferation.

Menin which is encoded by the Multiple Endocrine Neoplasia type 1 (MEN1)gene is expressed ubiquitously and is predominantly localized in thenucleus. It has been shown to interact with numerous proteins and is,therefore, involved in a variety of cellular processes. The bestunderstood function of menin is its role as an oncogenic cofactor of MLLfusion proteins. Menin interacts with two motifs within the N-terminalfragment of MLL that is retained in all fusion proteins, MBM1(menin-binding motif 1) and MBM2 (Thiel et al., Bioessays 2012. 34,771-80). Menin/MLL interaction leads to the formation of a newinteraction surface for lens epithelium-derived growth factor (LEDGF).Although MLL directly binds to LEDGF, menin is obligatory for the stableinteraction between MLL and LEDGF and the gene specific chromatinrecruitment of the MLL complex via the PWWP domain of LEDGF (Cermakovaet al., Cancer Res 2014. 15, 5139-51; Yokoyama & Cleary, Cancer Cell2008. 8, 36-46). Furthermore, numerous genetic studies have shown thatmenin is strictly required for oncogenic transformation by MLL fusionproteins suggesting the menin/MLL interaction as an attractivetherapeutic target. For example, conditional deletion of Men1 preventsleukomogenesis in bone marrow progenitor cells ectopically expressingMLL fusions (Chen et al., Proc Natl Acad Sci 2006. 103, 1018-23).Similarly, genetic disruption of menin/MLL fusion interaction byloss-of-function mutations abrogates the oncogenic properties of the MLLfusion proteins, blocks the development of leukemia in vivo and releasesthe differentiation block of MLL-transformed leukemic blasts. Thesestudies also showed that menin is required for the maintenance of HOXgene expression by MLL fusion proteins (Yokoyama et al., Cell 2005. 123,207-18). In addition, small molecule inhibitors of menin/MLL interactionhave been developed suggesting druggability of this protein/proteininteraction and have also demonstrated efficacy in preclinical models ofAML (Borkin et al., Cancer Cell 2015. 27, 589-602; Cierpicki andGrembecka, Future Med Chem 2014. 6, 447-462). Together with theobservation that menin is not a requisite cofactor of MLL1 during normalhematopoiesis (Li et al., Blood 2013. 122, 2039-2046), these datavalidate the disruption of menin/MLL interaction as a promising newtherapeutic approach for the treatment of MLL rearranged leukemia andother cancers with an active HOX/MEIS1 gene signature. For example, aninternal partial tandem duplication (PTD) within the 5′region of the MLLgene represents another major aberration that is found predominantly inde novo and secondary AML as well as myeloid dysplasia syndromes.Although the molecular mechanism and the biological function of MLL-PTDis not well understood, new therapeutic targeting strategies affectingthe menin/MLL interaction might also prove effective in the treatment ofMLL-PTD-related leukemias. Furthermore, castration-resistant prostatecancer has been shown to be dependent on the menin/MLL interaction(Malik et al., Nat Med 2015. 21, 344-52).

Several references describe inhibitors targeting the menin-MLLinteraction: WO2011029054, J Med Chem 2016, 59, 892-913 describes thepreparation of thienopyrimidine and benzodiazepine derivatives;WO2014164543 describes thienopyrimidine and thienopyridine derivatives;Nature Chemical Biology March 2012, 8, 277-284 and Ren, J.; et al.Bioorg Med Chem Lett (2016),http://dx.doi.org/10.1016/j.bmcl.2016.07.074 describe thienopyrimidinederivatives; J Med Chem 2014, 57, 1543-1556 describes hydroxy- andaminomethylpiperidine derivatives; and Future Med Chem 2014, 6, 447-462reviews small molecule and peptidomimetic compounds. WO2017112768describes inhibitors of the menin-MLL interaction.

DESCRIPTION OF THE INVENTION

The present invention concerns novel compounds of Formula (I):

and the tautomers and the stereoisomeric forms thereof, whereinR¹ is selected from the group consisting of CH₃, CH₂F, CHF₂, and CF₃;R² is selected from the group consisting of hydrogen and CH₃;L¹ represents a 7- to 10-membered saturated spiroheterobicyclic systemcontaining one or two N-atoms provided that it is N-linked to thethienopyrimidinyl heterocycle; and--L²-R³ is selected from (a), (b), (c), (d), (e), (f) or (g), wherein

-   (a) L² is selected from the group consisting of >SO₂,    >CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—; wherein    -   (i) when L² is linked to a carbon atom of L¹, then R^(4a) and R⁵        are each independently selected from the group consisting of        hydrogen; —OR⁶; —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and        C-linked 4- to 7-membered non-aromatic heterocyclyl containing        at least one nitrogen, oxygen or sulfur atom;    -   (ii) when L² is linked to a nitrogen atom of L¹, then R⁴ is        selected from the group consisting of hydrogen;        —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a        substituent selected from the group consisting of fluoro, —CN,        —OR⁸, and —NR^(9a)R^(9b); and C-linked 4- to 7-membered        non-aromatic heterocyclyl containing at least one nitrogen,        oxygen or sulfur atom; R⁵ is selected from the group consisting        of hydrogen; —OR⁶; —NR^(7a)R^(7b); —C(═O)N^(7a)R^(7b); C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and        C-linked 4- to 7-membered non-aromatic heterocyclyl containing        at least one nitrogen, oxygen or sulfur atom;    -   R^(4b) is selected from the group consisting of hydrogen and        methyl; or R^(4a) and R^(4b) together with the carbon atom to        which they are attached form a C₃₋₅cycloalkyl or a C-linked 4-        to 6-membered heterocyclyl containing an oxygen atom; wherein        R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each independently        selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, and —C(═O)NR^(10a)R^(10b); and        C₂₋₄alkyl substituted with a substituent selected from the group        consisting of —OR¹¹ and —NR^(10a)R^(10b); wherein    -   R^(10a), R^(10b) and R¹¹ are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; and    -   R³ is selected from the group consisting of Ar, Het¹, Het², and        a 7- to 10-membered saturated spirocarbobicyclic system; or-   (b) L² is selected from the group consisting of >CR^(4c)R^(4d) and    —CHR^(4c)CHR^(5a)—, wherein R^(4c), R^(4d), and R^(5a) are each    independently selected from the group consisting of hydrogen and    C₁₋₄alkyl; and    -   R³ is selected from the group consisting of

wherein R^(12a), R^(12b), and R^(12c) are each independently selectedfrom the group consisting of C₁₋₆alkyl optionally substituted with a —OHor a —NH₂ substituent; and —OC₁₋₆alkyl; or

-   (c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or    three fluoro substituents; or-   (d) L² is O and R³ is selected from the group consisting of    C₃₋₆alkyl optionally substituted with one, two or three fluoro    substituents; Ar; Het¹; Het²; a 7- to 10-membered saturated    spirocarbobicyclic system; —CH₂—Ar; —CH₂-Het¹; —CH₂-Het²; and    —CH₂-(a 7- to 10-membered saturated spirocarbobicyclic system); when    L² is linked to a carbon atom of L¹; or-   (e) --L²-R³ is —O—CHR⁵—R³ when L² is linked to a carbon atom of L¹,    wherein    -   R⁵ is selected from the group consisting of        —C(═O)NR^(13a)R^(13b); C₁₋₄alkyl optionally substituted with a        substituent selected from the group consisting of fluoro, —OR¹⁴,        and —NR^(15a)R^(15b); and C-linked 4- to 7-membered non-aromatic        heterocyclyl containing at least one nitrogen, oxygen or sulfur        atom; wherein    -   R^(13a), R^(13b), R¹⁴, R^(15a) and R^(15b) are each        independently selected from the group consisting of hydrogen;        C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro and —C(═O)NR^(16a)R^(16b);        and C₂₋₄alkyl substituted with a substituent selected from the        group consisting of —OR¹⁷ and —NR^(16a)R^(16b); wherein    -   R^(16a), R^(16b) and R¹⁷ are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; and    -   R³ is selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with one, two, or three fluoro        substituents; —CN; Ar, Het¹; Het²; and a 7- to 10-membered        saturated spirocarbobicyclic system; or-   (f) --L²-R³ is

wherein

-   -   R¹⁸ is selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with a fluoro or a —CN substituent; and        C₂₋₄alkyl substituted with a substituent selected from the group        consisting of —OR¹⁹ and —NR^(20a)R^(20b); wherein    -   R¹⁹, R^(20a) and R^(20b) are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl optionally        substituted with a substituent selected from the group        consisting of fluoro, —CN, and —C(═O)NR^(21a)R^(21b); C₂₋₄alkyl        substituted with a substituent selected from the group        consisting of —OR²² and —NR^(21a)R^(21b); and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; wherein    -   R^(21a), R^(21b) and R²² are each independently selected from        the group consisting of hydrogen and C₁₋₄alkyl; and    -   R^(18a) is selected from the group consisting of hydrogen,        fluoro and C₁₋₄alkyl;    -   R^(18b) is selected from the group consisting of fluoro,        —OC₁₋₄alkyl, and C₁₋₄alkyl optionally substituted with 1, 2 or 3        fluoro substituents; or    -   R^(18a) and R^(18b) are bound to the same carbon atom and        together form a C₃₋₅cycloalkyl or a C-linked 4- to 6-membered        heterocyclyl containing an oxygen atom; or

-   (g) --L²-R³ is

and whereinAr is phenyl or naphthyl, each of which may be optionally substitutedwith one, two, or three substituents each independently selected fromthe group consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), andC₁₋₄alkyl optionally substituted with a substituent selected from thegroup consisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b);Het¹ is a monocyclic heteroaryl selected from the group consisting ofpyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl,thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl,isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclic heteroarylselected from the group consisting of imidazothiazolyl,imidazoimidazolyl, benzofuranyl, benzothiophenyl, benzimidazolyl,benzoxazolyl, isobenzoxazolyl, benzisoxazolyl, benzothiazolyl,benzisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolizinyl,indolinyl, isoindolinyl, indazolyl, pyrazolopyridinyl,pyrazolopyrimidinyl, imidazopyridinyl, imidazopyrazinyl,imidazopyridazinyl; each of which may be optionally substituted withone, two, or three substituents each independently selected from thegroup consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b); andHet² is a non-aromatic heterocyclyl optionally substituted with one,two, or three substituents each independently selected from the groupconsisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b);whereinR²⁴, R^(25a), R^(25b), R²⁶, R^(27a), and R^(27b) are each independentlyselected from the group consisting of hydrogen; C₁₋₄alkyl optionallysubstituted with a substituent selected from the group consisting offluoro and —C(═O)NR^(28a)R^(28b); and C₂₋₄alkyl substituted with asubstituent selected from the group consisting of —OR²⁹ and—NR^(28a)R^(28b); wherein R^(28a), R^(28b) and R²⁹ are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyl;and C-linked 4- to 7-membered non-aromatic heterocyclyl containing atleast one nitrogen, oxygen or sulfur atom;and the pharmaceutically acceptable salts and the solvates thereof,provided that the following compounds, and pharmaceutically acceptableaddition salts, and solvates thereof are excluded:

It is to be understood that the above proviso applies to all embodimentsof the present invention described hereinafter.

The present invention also relates to a pharmaceutical compositioncomprising a therapeutically effective amount of a compound of Formula(I), a pharmaceutically acceptable salt, or a solvate thereof, and apharmaceutically acceptable carrier or excipient.

Additionally, the invention relates to a compound of Formula (I), apharmaceutically acceptable salt, or a solvate thereof, for use as amedicament, and to a compound of Formula (I), a pharmaceuticallyacceptable salt, or a solvate thereof, for use in the treatment or inthe prevention of cancer, myelodysplastic syndrome (MDS) and diabetes.

In a particular embodiment, the invention relates to a compound ofFormula (I), a pharmaceutically acceptable salt, or a solvate thereof,for use in the treatment or in the prevention of cancer.

In a specific embodiment said cancer is selected from leukemias, myelomaor a solid tumor cancer (e.g. prostate cancer, lung cancer, breastcancer, pancreatic cancer, colon cancer, liver cancer, melanoma andglioblastoma, etc.). In some embodiments, the leukemias include acuteleukemias, chronic leukemias, myeloid leukemias, myelogeneous leukemias,lymphoblastic leukemias, lymphocytic leukemias, Acute myelogeneousleukemias (AML), Chronic myelogenous leukemias (CML), Acutelymphoblastic leukemias (ALL), Chronic lymphocytic leukemias (CLL), Tcell prolymphocytic leukemias (T-PLL), Large granular lymphocyticleukemia, Hairy cell leukemia (HCL), MLL-rearranged leukemias, MLL-PTDleukemias, MLL amplified leukemias, MLL-positive leukemias, leukemiasexhibiting HOX/MEIS1 gene expression signatures etc.

The invention also relates to the use of a compound of Formula (I), apharmaceutically acceptable salt, or a solvate thereof, in combinationwith an additional pharmaceutical agent for use in the treatment orprevention of cancer, myelodysplastic syndrome (MDS) and diabetes.

Furthermore, the invention relates to a process for preparing apharmaceutical composition according to the invention, characterized inthat a pharmaceutically acceptable carrier is intimately mixed with atherapeutically effective amount of a compound of Formula (I), apharmaceutically acceptable salt, or a solvate thereof.

The invention also relates to a product comprising a compound of Formula(I), a pharmaceutically acceptable salt, or a solvate thereof, and anadditional pharmaceutical agent, as a combined preparation forsimultaneous, separate or sequential use in the treatment or preventionof cancer, myelodysplastic syndrome (MDS) and diabetes.

Additionally, the invention relates to a method of treating orpreventing a cell proliferative disease in a warm-blooded animal whichcomprises administering to the said animal an effective amount of acompound of Formula (I), a pharmaceutically acceptable salt, or asolvate thereof, as defined herein, or a pharmaceutical composition orcombination as defined herein.

DETAILED DESCRIPTION OF THE INVENTION

The term ‘halo’ or ‘halogen’ as used herein represents fluoro, chloro,bromo and iodo.

The prefix ‘C_(x-y)’ (where x and y are integers) as used herein refersto the number of carbon atoms in a given group. Thus, a C₁₋₄alkyl groupcontains from 1 to 6 carbon atoms, a C₃₋₆cycloalkyl group contains from3 to 6 carbon atoms, and so on.

The term ‘C₁₋₄alkyl’ as used herein as a group or part of a grouprepresents a straight or branched chain saturated hydrocarbon radicalhaving from 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl,isopropyl, n-butyl, s-butyl, t-butyl and the like.

The term ‘C₂₋₄alkyl’ as used herein as a group or part of a grouprepresents a straight or branched chain saturated hydrocarbon radicalhaving from 2 to 4 carbon atoms, such as ethyl, n-propyl, isopropyl,n-butyl, s-butyl, t-butyl and the like.

The term ‘C₁₋₄alkyl’ as used herein as a group or part of a grouprepresents a straight or branched chain saturated hydrocarbon radicalhaving from 1 to 6 carbon atoms such as the groups defined for C₁₋₄alkyland n-pentyl, n-hexyl, 2-methylbutyl and the like.

The term ‘C₃₋₆alkyl’ as used herein as a group or part of a grouprepresents a straight or branched chain saturated hydrocarbon radicalhaving from 3 to 6 carbon atoms such as n-propyl, isopropyl, n-butyl,s-butyl, t-butyl, n-pentyl, n-hexyl, 2-methylbutyl and the like.

The term ‘C₃₋₅cycloalkyl’ as used herein as a group or part of a groupdefines a saturated, cyclic hydrocarbon radical having from 3 to 5carbon atoms, such as cyclopropyl, cyclobutyl and cyclopentyl.

The term ‘spirobicyclic’ as used herein as group or part of a grouprepresents cyclic systems wherein two cycles are joined at a singleatom. Examples of these systems are 7- to 10-membered saturatedspiroheterobicyclic systems containing one or two N-atoms, wherein oneof the nitrogen atoms is always linked to the thienopyrimidinylheterocycle in the compounds of Formula (I) as defined herein. Suchspirocyclic systems include, but are not limited to systems resultingfrom the combination of e.g. piperidine, pyrrolidine, azetidine, andcyclobutane rings. Examples of such systems include, but are not limitedto (a), (b), (c), (d), (e), (f) and (g) below and the like

wherein a represents the position of linkage to the thienopyrimidinylheterocycle. The skilled person will understand that in these particularexamples, the options for --L²-R³ defined herein when L² is linked to anitrogen atom of L¹, apply to examples (a)-(f); while the options for--L²-R³ defined herein when L² is linked to a carbon atom of L¹, applyto example (g).

Examples of 7- to 10-membered saturated spirocarbobicyclic systemsinclude, but are not limited to

and the like.

In general, whenever the term ‘substituted’ is used in the presentinvention, it is meant, unless otherwise indicated or clear from thecontext, to indicate that one or more hydrogens, in particular from 1 to4 hydrogens, more in particular from 1 to 3 hydrogens, preferably 1 or 2hydrogens, more preferably 1 hydrogen, on the atom or radical indicatedin the expression using ‘substituted’ are replaced with a selection fromthe indicated group, provided that the normal valency is not exceeded,and that the substitution results in a chemically stable compound, i.e.a compound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture.

Combinations of substituents and/or variables are permissible only ifsuch combinations result in chemically stable compounds. ‘Stablecompound’ is meant to indicate a compound that is sufficiently robust tosurvive isolation to a useful degree of purity from a reaction mixture.

The skilled person will understand that when an atom or radical issubstituted with ‘a substituent’, it is meant that the atom or radicalreferred to is substituted with one substituent selected from theindicated group.

The skilled person will understand that the term ‘optionallysubstituted’ means that the atom or radical indicated in the expressionusing ‘optionally substituted’ may or may not be substituted (this meanssubstituted or unsubstituted respectively).

When two or more substituents are present on a moiety they may, wherepossible and unless otherwise indicated or clear from the context,replace hydrogens on the same atom or they may replace hydrogen atoms ondifferent atoms in the moiety.

It will be clear for the skilled person that, unless otherwise isindicated or is clear from the context, a substituent on a heterocyclylgroup may replace any hydrogen atom on a ring carbon atom or on a ringheteroatom.

Within the context of this invention ‘saturated’ means ‘fullysaturated’, if not otherwise specified.

A ‘non-aromatic group’ embraces unsaturated ring systems withoutaromatic character, partially saturated and fully saturated carbocyclicand heterocyclic ring systems. The term ‘partially saturated’ refers torings wherein the ring structure(s) contain(s) at least one multiplebond e.g. a C═C, N═C bond. The term ‘fully saturated’ refers to ringswhere there are no multiple bonds between ring atoms. Thus, a‘non-aromatic heterocyclyl’ is a non-aromatic monocyclic or bicyclicsystem, unless otherwise specified, having for example, 3 to 12 ringmembers, more usually 5 to 10 ring members. Examples of monocyclicgroups are groups containing 4 to 7 ring members, more usually, 5 or 6ring members. Examples of bicyclic groups are those containing 8 to 12,more usually 9 or 10 ring members.

Non-limiting examples of monocyclic heterocyclyl systems containing atleast one heteroatom selected from nitrogen, oxygen or sulfur (N, O, S)include, but are not limited to 4- to 7-membered heterocyclyl systemssuch as azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl,piperidinyl, piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl,morpholinyl, thiomorpholinyl, and tetrahydro-2H-thiopyranyl 1,1-dioxide,in particular azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl,piperidinyl, piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl,morpholinyl, and thiomorpholiny. Non-limiting examples of bicyclicheterocyclyl systems containing at least one heteroatom selected fromnitrogen, oxygen or sulfur (N, O, S) include, but are not limited tooctahydro-1H-indolyl, indolinyl

Unless otherwise specified, each can be bound to the remainder of themolecule of Formula (I) through any available ring carbon atom(C-linked) or nitrogen atom (N-linked), and may optionally besubstituted, where possible, on carbon and/or nitrogen atoms accordingto the embodiments.

Examples of a C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen atom include, but are not limited to,azetidinyl, pyrrolidinyl and piperidinyl, bound to the rest of themolecule through an available carbon atom.

The term ‘C-linked 4- to 6-membered heterocyclyl containing an oxygenatom’ as used herein alone or as part of another group, defines asaturated, cyclic hydrocarbon radical containing an oxygen atom havingfrom 4 to 6 ring members, such as oxetanyl, tetrahydrofuranyl, andtetrahydropyranyl.

Whenever substituents are represented by chemical structure, ‘---’represents the bond of attachment to the remainder of the molecule ofFormula (I).

Lines (such as ‘---’) drawn into ring systems indicate that the bond maybe attached to any of the suitable ring atoms.

Het¹ and Het² may be attached to the remainder of the molecule ofFormula (I) through any available ring carbon or nitrogen atom asappropriate, if not otherwise specified.

It will be clear that a saturated cyclic moiety may, where possible,have substituents on both carbon and N-atoms, unless otherwise isindicated or is clear from the context.

It will be clear that when L² is >SO₂, this is equivalent to L² is—SO₂—. It will be clear that when L² is >CR^(4a)R^(4b), this isequivalent to L is

For example, in compound 1, L² is >CR^(4a)R^(4b) wherein both R^(4a) andR^(4b) are hydrogen.

Similar, it will be clear that when L² is >CR^(4c)R^(4d), this isequivalent to L is

When any variable occurs more than one time in any constituent, eachdefinition is independent.

When any variable occurs more than one time in any formula (e.g. Formula(I)), each definition is independent.

The term ‘subject’ as used herein, refers to an animal, preferably amammal (e.g. cat, dog, primate or human), more preferably a human, whois or has been the object of treatment, observation or experiment.

The term ‘therapeutically effective amount’ as used herein, means thatamount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue system, animal or humanthat is being sought by a researcher, veterinarian, medicinal doctor orother clinician, which includes alleviation or reversal of the symptomsof the disease or disorder being treated.

The term ‘composition’ is intended to encompass a product comprising thespecified ingredients in the specified amounts, as well as any productwhich results, directly or indirectly, from combinations of thespecified ingredients in the specified amounts.

The term ‘treatment’, as used herein, is intended to refer to allprocesses wherein there may be a slowing, interrupting, arresting orstopping of the progression of a disease, but does not necessarilyindicate a total elimination of all symptoms.

The term ‘compound(s) of the (present) invention’ or ‘compound(s)according to the (present) invention’ as used herein, is meant toinclude the compounds of Formula (I) and the pharmaceutically acceptablesalts, and the solvates thereof.

As used herein, any chemical formula with bonds shown only as solidlines and not as solid wedged or hashed wedged bonds, or otherwiseindicated as having a particular configuration (e.g. R, S) around one ormore atoms, contemplates each possible stereoisomer, or mixture of twoor more stereoisomers.

Hereinbefore and hereinafter, the term ‘compound(s) of Formula (I)’ ismeant to include the tautomers thereof and the stereoisomeric formsthereof.

The terms ‘stereoisomers’, ‘stereoisomeric forms’ or ‘stereochemicallyisomeric forms’ hereinbefore or hereinafter are used interchangeably.

The invention includes all stereoisomers of the compounds of theinvention either as a pure stereoisomer or as a mixture of two or morestereoisomers.

Enantiomers are stereoisomers that are non-superimposable mirror imagesof each other. A 1:1 mixture of a pair of enantiomers is a racemate orracemic mixture.

Atropisomers (or atropoisomers) are stereoisomers which have aparticular spatial configuration, resulting from a restricted rotationabout a single bond, due to large steric hindrance. All atropisomericforms of the compounds of Formula (I) are intended to be included withinthe scope of the present invention.

Diastereomers (or diastereoisomers) are stereoisomers that are notenantiomers, i.e. they are not related as mirror images. If a compoundcontains a double bond, the substituents may be in the E or the Zconfiguration.

Substituents on bivalent cyclic saturated or partially saturatedradicals may have either the cis- or trans-configuration; for example ifa compound contains a disubstituted cycloalkyl group, the substituentsmay be in the cis or trans configuration.

Therefore, the invention includes enantiomers, atropisomers,diastereomers, racemates, E isomers, Z isomers, cis isomers, transisomers and mixtures thereof, whenever chemically possible.

The meaning of all those terms, i.e. enantiomers, atropisomers,diastereomers, racemates, E isomers, Z isomers, cis isomers, transisomers and mixtures thereof are known to the skilled person.

The absolute configuration is specified according to theCahn-Ingold-Prelog system. The configuration at an asymmetric atom isspecified by either R or S. Resolved stereoisomers whose absoluteconfiguration is not known can be designated by (+) or (−) depending onthe direction in which they rotate plane polarized light. For instance,resolved enantiomers whose absolute configuration is not known can bedesignated by (+) or (−) depending on the direction in which they rotateplane polarized light.

When a specific stereoisomer is identified, this means that saidstereoisomer is substantially free, i.e. associated with less than 50%,preferably less than 20%, more preferably less than 10%, even morepreferably less than 5%, in particular less than 2% and most preferablyless than 1%, of the other stereoisomers. Thus, when a compound ofFormula (I) is for instance specified as (R), this means that thecompound is substantially free of the (S) isomer; when a compound ofFormula (I) is for instance specified as E, this means that the compoundis substantially free of the Z isomer; when a compound of Formula (I) isfor instance specified as cis, this means that the compound issubstantially free of the trans isomer.

Some of the compounds according to Formula (I) may also exist in theirtautomeric form. Such forms in so far as they may exist, although notexplicitly indicated in the above Formula (I) are intended to beincluded within the scope of the present invention. It follows that asingle compound may exist in both stereoisomeric and tautomeric form.

Pharmaceutically acceptable salts include acid addition salts and baseaddition salts. Such salts may be formed by conventional means, forexample by reaction of a free acid or a free base form with one or moreequivalents of an appropriate base or acid, optionally in a solvent, orin a medium in which the salt is insoluble, followed by removal of saidsolvent, or said medium, using standard techniques (e.g. in vacuo, byfreeze-drying or by filtration). Salts may also be prepared byexchanging a counter-ion of a compound of the invention in the form of asalt with another counter-ion, for example using a suitable ion exchangeresin.

The pharmaceutically acceptable salts as mentioned hereinabove orhereinafter are meant to comprise the therapeutically active non-toxicacid and base salt forms which the compounds of Formula (I) and solvatesthereof, are able to form.

Appropriate acids comprise, for example, inorganic acids such ashydrohalic acids, e.g. hydrochloric or hydrobromic acid, sulfuric,nitric, phosphoric and the like acids; or organic acids such as, forexample, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e.ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic,fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic,benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic,p-aminosalicylic, pamoic and the like acids. Conversely said salt formscan be converted by treatment with an appropriate base into the freebase form.

The compounds of Formula (I) and solvates thereof containing an acidicproton may also be converted into their non-toxic metal or amine saltforms by treatment with appropriate organic and inorganic bases.

Appropriate base salt forms comprise, for example, the ammonium salts,the alkali and earth alkaline metal salts, e.g. the lithium, sodium,potassium, cesium, magnesium, calcium salts and the like, salts withorganic bases, e.g. primary, secondary and tertiary aliphatic andaromatic amines such as methylamine, ethylamine, propylamine,isopropylamine, the four butylamine isomers, dimethylamine,diethylamine, diethanolamine, dipropylamine, diisopropylamine,di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine,triethylamine, tripropylamine, quinuclidine, pyridine, quinoline andisoquinoline; the benzathine, N-methyl-D-glucamine, hydrabamine salts,and salts with amino acids such as, for example, arginine, lysine andthe like. Conversely the salt form can be converted by treatment withacid into the free acid form.

The term solvate comprises the solvent addition forms as well as thesalts thereof, which the compounds of Formula (I) are able to form.Examples of such solvent addition forms are e.g. hydrates, alcoholatesand the like.

The compounds of the invention as prepared in the processes describedbelow may be synthesized in the form of mixtures of enantiomers, inparticular racemic mixtures of enantiomers, that can be separated fromone another following art-known resolution procedures. A manner ofseparating the enantiomeric forms of the compounds of Formula (I), andpharmaceutically acceptable salts, and solvates thereof, involves liquidchromatography using a chiral stationary phase. Said purestereochemically isomeric forms may also be derived from thecorresponding pure stereochemically isomeric forms of the appropriatestarting materials, provided that the reaction occursstereospecifically. Preferably if a specific stereoisomer is desired,said compound would be synthesized by stereospecific methods ofpreparation. These methods will advantageously employ enantiomericallypure starting materials.

The present invention also embraces isotopically-labeled compounds ofthe present invention which are identical to those recited herein, butfor the fact that one or more atoms are replaced by an atom having anatomic mass or mass number different from the atomic mass or mass numberusually found in nature (or the most abundant one found in nature).

All isotopes and isotopic mixtures of any particular atom or element asspecified herein are contemplated within the scope of the compounds ofthe invention, either naturally occurring or synthetically produced,either with natural abundance or in an isotopically enriched form.Exemplary isotopes that can be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorus, sulfur, fluorine, chlorine and iodine, such as ²H, ³H, ¹¹C,¹³C, ¹⁴C, ¹³N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P, ³³P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²²I, ¹²³I,¹²⁵I, ¹³¹I, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br and ⁸²Br. Preferably, the radioactiveisotope is selected from the group of ²H, ³H, ¹¹C and ¹⁸F. Morepreferably, the radioactive isotope is ²H. In particular, deuteratedcompounds are intended to be included within the scope of the presentinvention.

Certain isotopically-labeled compounds of the present invention (e.g.,those labeled with ³H and ¹⁴C) may be useful for example in substratetissue distribution assays. Tritiated (³H) and carbon-14 (¹⁴C) isotopesare useful for their ease of preparation and detectability. Further,substitution with heavier isotopes such as deuterium (i.e., ²H mayafford certain therapeutic advantages resulting from greater metabolicstability (e.g., increased in vivo half-life or reduced dosagerequirements) and hence may be preferred in some circumstances. Thus, ina particular embodiment of the present invention, R² is selected fromhydrogen or deuterium, in particular deuterium. In another embodiment,L² can be >C(²H)₂. Positron emitting isotopes such as ¹⁵O, ¹³N, ¹¹C and¹⁸F are useful for positron emission tomography (PET) studies. PETimaging in cancer finds utility in helping locate and identify tumours,stage the disease and determine suitable treatment. Human cancer cellsoverexpress many receptors or proteins that are potentialdisease-specific molecular targets. Radiolabelled tracers that bind withhigh affinity and specificity to such receptors or proteins on tumourcells have great potential for diagnostic imaging and targetedradionuclide therapy (Charron, Carlie L. et al. Tetrahedron Lett. 2016,57(37), 4119-4127). Additionally, target-specific PET radiotracers maybe used as biomarkers to examine and evaluate pathology, by for example,measuring target expression and treatment response (Austin R. et al.Cancer Letters (2016), doi: 10.1016/j.canlet.2016.05.008).

The present invention relates in particular to compounds of Formula (I)and the pharmaceutically acceptable salts, and the solvates thereof, orany subgroup thereof as mentioned in any of the embodiments, whereinalso the following compound and pharmaceutically acceptable additionsalts, and solvates thereof, are excluded:

The present invention relates in particular to compounds of Formula (I)and the pharmaceutically acceptable salts, and the solvates thereof, orany subgroup thereof as mentioned in any of the embodiments, wherein theintermediates and compounds described in WO2017/112768, in as far asthey are covered by the present invention, are excluded.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

R¹ is CF₃;R² is selected from the group consisting of hydrogen and CH₃;L¹ represents a 7- to 10-membered saturated spiroheterobicyclic systemcontaining one or two N-atoms provided that it is N-linked to thethienopyrimidinyl heterocycle; and--L²-R³ is selected from (a), (b), (c), (d), (f) or (g), wherein

-   (a) L² is selected from the group consisting of >CR^(4a)R^(4b), and    —CHR^(4a)CHR⁵—; wherein    -   L² is linked to a nitrogen atom of L¹; R^(4a) is selected from        the group consisting of hydrogen; —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of —OR⁸, and —NR^(9a)R^(9b); and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen atom;    -   R⁵ is selected from the group consisting of hydrogen; —OR⁶; and        C₁₋₄alkyl;    -   R^(4b) is selected from the group consisting of hydrogen and        methyl; or    -   R^(4a) and R^(4b) together with the carbon atom to which they        are attached form a C₃₋₅cycloalkyl or a C-linked 4- to        6-membered heterocyclyl containing an oxygen atom; wherein    -   R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each independently        selected from the group consisting of hydrogen; and C₂₋₄alkyl        substituted with a substituent selected from the group        consisting of —OR¹¹ and —NR^(10a)R^(10b); wherein    -   R^(10a), R^(10b) and R¹¹ are each independently selected from        the group consisting of hydrogen; and C₁₋₄alkyl; and    -   R³ is selected from the group consisting of Ar, Het¹, Het², and        a 7- to 10-membered saturated spirocarbobicyclic system; or    -   (b) L² is >CR^(4c)R^(4d), wherein R^(4c) and R^(4d) are        hydrogen; and    -   R³ is

wherein R^(12a), R^(12b), and R^(12c) are C₁₋₆alkyl 1; or

-   (c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or    three fluoro substituents; or-   (d) L² is O and R³ is —CH₂—Ar; or-   (f) --L²-R³ is

wherein

-   -   R¹⁸ is selected from the group consisting of hydrogen; and        C₁₋₄alkyl;    -   R^(18a) is selected from the group consisting of hydrogen, and        fluoro;    -   R^(18b) is selected from the group consisting of fluoro,        —OC₁₋₄alkyl, and C₁₋₄alkyl optionally substituted with 1, 2 or 3        fluoro substituents; or    -   R^(18a) and R^(18b) are bound to the same carbon atom and        together form a C₃₋₅cycloalkyl; or

-   (g) --L²-R³ is

and whereinAr is phenyl which may be optionally substituted with one, two, or threesubstituents each independently selected from the group consisting ofhalo, —OR²⁴, and C₁₋₄alkyl optionally substituted with —OR²⁶;Het¹ is a monocyclic heteroaryl selected from the group consisting ofpyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl,pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl, and isoxazolyl;or a bicyclic heteroaryl selected from the group consisting of indolyl,imidazopyridinyl; each of which may be optionally substituted with one,two, or three substituents each independently selected from the groupconsisting of halo, —CN, —OR²⁴, and C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of —CN, —OR²⁶, and—NR^(27a)R^(27b); andHet² is a non-aromatic heterocyclyl optionally substituted with one,two, or three substituents each independently selected from the groupconsisting of halo, —CN, and C₁₋₄alkyl optionally substituted with—OR²⁶;whereinR²⁴, R²⁶, R^(27a), and R^(27b) are each independently selected from thegroup consisting of hydrogen; C₁₋₄alkyl; and C₂₋₄alkyl substituted with—NR^(28a)R^(28b); whereinR^(28a) and R^(28b) are hydrogen;and the pharmaceutically acceptable salts and the solvates thereof.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

-   (a) L² is selected from the group consisting of >SO₂,    >CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—; wherein    -   (i) when L² is linked to a carbon atom of L¹, then R^(4a) and R⁵        are each independently selected from the group consisting of        hydrogen; —OR⁶; —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and        C-linked 4- to 7-membered non-aromatic heterocyclyl containing        at least one nitrogen, oxygen or sulfur atom;    -   (ii) when L² is linked to a nitrogen atom of L¹, then R^(4a) is        selected from the group consisting of hydrogen;        —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a        substituent selected from the group consisting of fluoro, —CN,        —OR⁸, and —NR^(9a)R^(9b); and C-linked 4- to 7-membered        non-aromatic heterocyclyl containing at least one nitrogen,        oxygen or sulfur atom;        -   R⁵ is selected from the group consisting of hydrogen; —OR⁶;            —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally            substituted with a substituent selected from the group            consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and            C-linked 4- to 7-membered non-aromatic heterocyclyl            containing at least one nitrogen, oxygen or sulfur atom;    -   R⁴ is selected from the group consisting of hydrogen and methyl;        or >CR^(4a)R^(4b) form a >C₃₋₅cycloalkanediyl or a >C-linked 4-        to 6-membered heterocyclediyl containing an oxygen atom; wherein    -   R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each independently        selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, and —C(═O)NR^(10a)R^(10b); and        C₂₋₄alkyl substituted with a substituent selected from the group        consisting of —OR¹¹ and —NR^(10a)R^(10b); wherein    -   R^(10a), R^(10b) and R¹¹ are each independently selected from        the group consisting of hydrogen; and C₁₋₄alkyl; and    -   R³ is selected from the group consisting of Ar, Het¹, Het², and        a 7- to 10-membered saturated spirocarbobicyclic system; or-   (b) L² is selected from the group consisting of >CR^(4c)R^(4d) and    —CHR^(4c)CHR^(5a)—, wherein R^(4c), R^(4d), and R^(5a) are each    independently selected from the group consisting of hydrogen and    C₁₋₄alkyl; and    -   R³ is selected from the group consisting of

wherein R^(12a), R^(12b), and R^(12c) are each independently selectedfrom the group consisting of C₁₋₆alkyl optionally substituted with a —OHor a —NH₂ substituent; or

-   (c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or    three fluoro substituents; or-   (d) L² is O and R³ is selected from the group consisting of Ar,    Het¹; —CH₂—Ar, —CH₂-Het¹, and —CH₂-(a 7- to 10-membered saturated    spirocarbobicyclic system); when L² is linked to a carbon atom of    L¹; or-   (e) --L²-R³ is selected from the group consisting of

wherein

-   -   R¹⁸ is hydrogen; or

-   (f) --L²-R³ is

and whereinAr is phenyl optionally substituted with one, two, or three substituentseach independently selected from the group consisting of halo, —CN, andC₁₋₄alkyl optionally substituted with a substituent selected from thegroup consisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b);Het¹ is a monocyclic heteroaryl selected from the group consisting ofpyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl,thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl,isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclic heteroarylselected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl;each of which may be optionally substituted with one, two, or threesubstituents each independently selected from the group consisting ofhalo, —CN, and C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); andHet² is a non-aromatic heterocyclyl selected from azetidinyl,pyrrolidinyl and piperidinyl;whereinR²⁶, R^(27a), and R^(27b) are each independently selected from the groupconsisting of hydrogen and C₁₋₄alkyl;and the pharmaceutically acceptable salts and the solvates thereof.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

R¹ is selected from the group consisting of CH₃, CH₂F, CHF₂, and CF₃;R² is selected from the group consisting of hydrogen and CH₃;L¹ represents a 7- to 10-membered saturated spiroheterobicyclic systemcontaining one or two N-atoms provided that it is N-linked to thethienopyrimidinyl heterocycle; and--L²-R³ is selected from (a), (b), (d), (e), or (f), wherein

-   (a) L² is selected from the group consisting of >SO₂,    >CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—; wherein    -   (i) when L² is linked to a carbon atom of L¹, then R^(4a) and R⁵        are each independently selected from the group consisting of        hydrogen; —OR⁶; —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and        C-linked 4- to 7-membered non-aromatic heterocyclyl containing        at least one nitrogen, oxygen or sulfur atom;    -   (ii) when L² is linked to a nitrogen atom of L¹, then R^(4a) is        selected from the group consisting of —C(═O)NR^(7a)R^(7b);        C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro, —CN, —OR⁸, and        —NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic        heterocyclyl containing at least one nitrogen, oxygen or sulfur        atom;        -   R⁵ is selected from the group consisting of hydrogen; —OR⁶;            —NR^(7a)R^(7b); —C(═O)N^(7a)R^(7b); C₁₋₄alkyl optionally            substituted with a substituent selected from the group            consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and            C-linked 4- to 7-membered non-aromatic heterocyclyl            containing at least one nitrogen, oxygen or sulfur atom;    -   R^(4b) is selected from the group consisting of hydrogen and        methyl; or R^(4a) and R^(4b) together with the carbon atom to        which they are attached form a C₃₋₅cycloalkyl or a C-linked 4-        to 6-membered heterocyclyl containing an oxygen atom; wherein        -   R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each            independently selected from the group consisting of            hydrogen; C₁₋₄alkyl optionally substituted with a            substituent selected from the group consisting of fluoro,            —CN, and —C(═O)NR^(10a)R^(10b); and C₂₋₄alkyl substituted            with a substituent selected from the group consisting of            —OR¹¹ and —NR^(10a)R^(10b); wherein    -   R^(10a), R^(10b) and R¹¹ are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; and    -   R³ is selected from the group consisting of Ar, Het¹, Het², and        a 7- to 10-membered saturated spirocarbobicyclic system; or-   (b) L² is selected from the group consisting of >CR^(4c)R^(4d) and    —CHR^(4c)CHR^(5a)—, wherein R^(4c), R^(4d), and R^(5a) are each    independently selected from the group consisting of hydrogen and    C₁₋₄alkyl; and    -   R³ is selected from the group consisting of

wherein R^(12a), R^(12b), and R^(12c) are each independently selectedfrom the group consisting of C₁₋₆alkyl optionally substituted with a —OHor a —NH₂ substituent; and —OC₁₋₄alkyl; or

-   (d) L² is O and R³ is selected from the group consisting of    C₃₋₆alkyl optionally substituted with one, two or three fluoro    substituents; Ar; Het¹; Het²; a 7- to 10-membered saturated    spirocarbobicyclic system; —CH₂—Ar; —CH₂-Het¹; —CH₂-Het²; and    —CH₂-(a 7- to 10-membered saturated spirocarbobicyclic system); when    L² is linked to a carbon atom of L¹; or-   (e) --L²-R³ is —O—CHR⁵—R³ when L² is linked to a carbon atom of L¹,    wherein    -   R⁵ is selected from the group consisting of        —C(═O)NR^(13a)R^(13b); C₁₋₄alkyl optionally substituted with a        substituent selected from the group consisting of fluoro, —OR¹⁴,        and —NR^(15a)R^(15b); and C-linked 4- to 7-membered non-aromatic        heterocyclyl containing at least one nitrogen, oxygen or sulfur        atom; wherein    -   R^(13a), R^(13b), R¹⁴, R^(15a) and R^(15b) are each        independently selected from the group consisting of hydrogen;        C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro and —C(═O)NR^(16a)R^(16b);        and C₂₋₄alkyl substituted with a substituent selected from the        group consisting of —OR¹⁷ and —NR^(16a)R^(16b); wherein    -   R^(16a), R^(16b) and R¹⁷ are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; and    -   R³ is selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with one, two, or three fluoro        substituents; —CN; Ar, Het¹; Het²; and a 7- to 10-membered        saturated spirocarbobicyclic system; or-   (f) --L²-R³ is

wherein

-   -   R¹⁸ is selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with a fluoro or a —CN substituent; and        C₂₋₄alkyl substituted with a substituent selected from the group        consisting of —OR¹⁹ and —NR^(20a)R^(20b); wherein    -   R¹⁹, R^(20a) and R^(20b) are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl optionally        substituted with a substituent selected from the group        consisting of fluoro, —CN, and —C(═O)NR^(21a)R^(21b); C₂₋₄alkyl        substituted with a substituent selected from the group        consisting of —OR²² and —NR^(21a)R^(21b); and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; wherein    -   R^(21a), R^(21b) and R²² are each independently selected from        the group consisting of hydrogen and C₁₋₄alkyl; and    -   R^(18a) is selected from the group consisting of hydrogen,        fluoro and C₁₋₄alkyl;    -   R^(18b) is selected from the group consisting of fluoro,        —OC₁₋₄alkyl, and C₁₋₄alkyl optionally substituted with 1, 2 or 3        fluoro substituents; or    -   R^(18a) and R^(18b) are bound to the same carbon atom and        together form a C₃₋₅cycloalkyl or a C-linked 4- to 6-membered        heterocyclyl containing an oxygen atom;    -   and wherein        Ar is phenyl or naphthyl, each of which may be optionally        substituted with one, two, or three substituents each        independently selected from the group consisting of halo, —CN,        —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted        with a substituent selected from the group consisting of fluoro,        —CN, —OR²⁶, —NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b);        Het¹ is a monocyclic heteroaryl selected from the group        consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl,        pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl,        4- or 5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl;        or a bicyclic heteroaryl selected from the group consisting of        imidazothiazolyl, imidazoimidazolyl, benzofuranyl,        benzothiophenyl, benzimidazolyl, benzoxazolyl, isobenzoxazolyl,        benzisoxazolyl, benzothiazolyl, benzisothiazolyl,        isobenzofuranyl, indolyl, isoindolyl, indolizinyl, indolinyl,        isoindolinyl, indazolyl, pyrazolopyridinyl, pyrazolopyrimidinyl,        imidazopyridinyl, imidazopyrazinyl, imidazopyridazinyl; each of        which may be optionally substituted with one, two, or three        substituents each independently selected from the group        consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyl        optionally substituted with a substituent selected from the        group consisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and        —C(═O)NR^(27a)R^(27b); and        Het² is a non-aromatic heterocyclyl optionally substituted with        one, two, or three substituents each independently selected from        the group consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and        C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro, —CN, —OR²⁶,        —NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b);        wherein        R²⁴, R^(25a), R^(25b), R²⁶, R^(27a), and R^(27b) are each        independently selected from the group consisting of hydrogen;        C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro and —C(═O)NR^(28a)R^(28b);        and C₂₋₄alkyl substituted with a substituent selected from the        group consisting of —OR²⁹ and —NR^(28a)R^(28b); wherein        R^(28a), R^(28b) and R²⁹ are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom;        and the pharmaceutically acceptable salts and the solvates        thereof.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

R¹ is CF₃;

-   (a) L² is >CR^(4a)R^(4b); wherein    -   R^(4a) is selected from the group consisting of hydrogen;        —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl; and C-linked 4- to 7-membered        non-aromatic heterocyclyl containing at least one nitrogen,        oxygen or sulfur atom; and    -   R^(4b) is selected from the group consisting of hydrogen and        methyl; wherein    -   R^(7a) and R^(7b) are each independently selected from the group        consisting of hydrogen; C₁₋₄alkyl; and C₂₋₄alkyl substituted        with a substituent selected from the group consisting of —OR¹¹        and —NR^(10a)R^(10b); wherein    -   R^(10a), R^(10b) and R¹¹ are each independently selected from        the group consisting of hydrogen and C₁₋₄alkyl; and    -   R³ is selected from the group consisting of Ar, Het¹, Het², and        a 7- to 10-membered saturated spirocarbobicyclic system; or-   (b) L² is >CR^(4c)R^(4d), wherein R^(4c) and R^(4d) are each    independently selected from the group consisting of hydrogen and    C₁₋₄alkyl; and    -   R³ is selected from the group consisting of

wherein R^(12a), R^(12b), and R^(12c) are each independently selectedfrom the group consisting of C₁₋₆alkyl optionally substituted with a—NH₂ substituent; or

-   (c) --L²-R³ is C₁₋₆-alkyl optionally substituted with one, two or    three fluoro substituents; or-   (d) L² is O and R³ is selected from the group consisting of Ar,    Het¹, —CH₂—Ar, —CH₂-Het¹, and —CH₂-(a 7- to 10-membered saturated    spirocarbobicyclic system); when L² is linked to a carbon atom of    L¹; or-   (e) --L²-R³ is selected from the group consisting of

wherein R¹⁸ is hydrogen; or

-   (f) --L²-R³ is

and whereinAr is phenyl optionally substituted with a halo substituent;Het¹ is a monocyclic heteroaryl selected from the group consisting ofpyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl,pyrazolyl, imidazolyl, and 4- or 5-thiazolyl; or a bicyclic heteroarylselected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl;each of which may be optionally substituted with one, two, or threesubstituents each independently selected from the group consisting ofhalo and C₁₋₄alkyl optionally substituted with a substituent selectedfrom the group consisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b); andHet² is a non-aromatic heterocyclyl selected from azetidinyl,pyrrolidinyl and piperidinyl;whereinR²⁶, R^(27a), and R^(27b) are each independently selected from the groupconsisting of hydrogen and C₁₋₄alkyl;and the pharmaceutically acceptable salts and the solvates thereof.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

R¹ is CF₃;L¹ represents a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (a), (b), (c), (d), (e), (f) and (g)

wherein a represents the position of linkage to the thienopyrimidinylheterocycle;

-   (a) L² is >CH₂; and R³ is selected from the group consisting of Ar,    Het¹, and a 7- to 10-membered saturated spirocarbobicyclic system;    or-   (b) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or    three fluoro substituents; and wherein    Ar is phenyl optionally substituted with a halo substituent; and    Het¹ is a monocyclic heteroaryl selected from the group consisting    of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl,    pyrazolyl, imidazolyl, and 4- or 5-thiazolyl; or a bicyclic    heteroaryl selected from imidazopyridinyl, in particular    imidazo[1,2-a]pyridinyl; each of which may be optionally substituted    with one or two substituents each independently selected from the    group consisting of halo and C₁₋₄alkyl;    and the pharmaceutically acceptable salts and the solvates thereof.

The present invention relates in particular to compounds of Formula (I)as defined herein, and the tautomers and the stereoisomeric formsthereof, wherein

R¹ is CF₃;R² is hydrogen;L¹ represents a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (a), (b), (c), (d), (e), (f) and (g)

wherein a represents the position of linkage to the thienopyrimidinylheterocycle;

-   (a) L² is >CH₂; and R³ is selected from the group consisting of Ar,    Het¹, and a 7- to 10-membered saturated spirocarbobicyclic system;    or-   (b) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or    three fluoro substituents; and wherein    Ar is phenyl optionally substituted with a halo substituent; and    Het¹ is a monocyclic heteroaryl selected from the group consisting    of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl,    pyrazolyl, and imidazolyl; or a bicyclic heteroaryl selected from    imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of    which may be optionally substituted with one or two substituents    each independently selected from the group consisting of halo and    C₁₋₄alkyl;    and the pharmaceutically acceptable salts and the solvates thereof.

Another embodiment of the present invention relates to those compoundsof Formula (I) and the pharmaceutically acceptable salts, and thesolvates thereof, or any subgroup thereof as mentioned in any of theother embodiments wherein one or more of the following restrictionsapply:

(a) R¹ is CF₃;(b) R² is hydrogen;(c) L¹ is a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (a), (b), (c), (d), (e), (f) and (g) as defined herein;(d) L¹ is a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (a), (b), (c), (d), (e), and (f) as defined herein;(e) L¹ is a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (c) and (e);(f) L² is >CH₂;(g) L² is >CH₂; and R³ is selected from the group consisting of Ar,Het¹, and a 7- to 10-membered saturated spirocarbobicyclic system;(h) --L²-R³ is selected from the group consisting of

wherein

-   -   R¹⁸ is selected from the group consisting of hydrogen; C₁₋₄alkyl        optionally substituted with a fluoro or —CN substituent; and        C₂₋₄alkyl substituted with a substituent selected from the group        consisting of —OR¹⁹ and —NR^(20a)R^(20b); wherein    -   R¹⁹, R^(20a) and R^(20b) are each independently selected from        the group consisting of hydrogen; C₁₋₄alkyl optionally        substituted with a substituent selected from the group        consisting of fluoro, —CN, and —C(═O)NR^(21a)R^(21b); C₂₋₄alkyl        substituted with a substituent selected from the group        consisting of —OR²² and —NR^(21a)R^(21b); and C-linked 4- to        7-membered non-aromatic heterocyclyl containing at least one        nitrogen, oxygen or sulfur atom; wherein    -   R^(21a), R^(21b) and R²² are each independently selected from        the group consisting of hydrogen and C₁₋₄alkyl;        (i) Ar is phenyl optionally substituted with one or two        independently selected halo substituents;        (j) Ar is phenyl optionally substituted with one halo        substituent;        (k) Ar is phenyl;        (l) Het¹ is a monocyclic heteroaryl selected from the group        consisting of pyrazolyl, imidazolyl, pyrrolyl, 4- or        5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl, and        pyrazinyl; or is a bicyclic heteroaryl selected from        imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of        which may be optionally substituted with one or two substituents        each independently selected from the group consisting of halo        and C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of fluoro, —CN, —OR²⁵,        —NR^(26a)R^(26b), and —C(═O)NR^(26a)R^(26b); wherein R²⁵,        R^(26a), and R^(26b) are each independently selected from the        group consisting of hydrogen and C₁₋₄alkyl;        (m) Het¹ is a monocyclic heteroaryl selected from the group        consisting of pyrazolyl, imidazolyl, pyrrolyl, 4- or        5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl, and        pyrazinyl; or is a bicyclic heteroaryl selected from        imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of        which may be optionally substituted with one or two substituents        each independently selected from the group consisting of halo        and C₁₋₄alkyl optionally substituted with a substituent selected        from the group consisting of —CN, —OR²⁵, —NR^(26a)R^(26b), and        —C(═O)NR^(26a)R^(26b); wherein R²⁵, R^(26a), and R^(26b) are        each independently selected from the group consisting of        hydrogen and C₁₋₄alkyl;        (n) Het¹ is a monocyclic heteroaryl selected from the group        consisting of pyrazolyl, imidazolyl, pyrrolyl, pyridazinyl, 4-,        5- or 6-pyrimidinyl, and pyrazinyl; or is a bicyclic heteroaryl        selected from imidazopyridinyl, in particular        imidazo[1,2-a]pyridinyl; each of which may be optionally        substituted with one or two substituents each independently        selected from the group consisting of halo and C₁₋₄alkyl;        (o) Het¹ is a monocyclic heteroaryl selected from the group        consisting of pyridazinyl, 4-, 5- or 6-pyrimidinyl, and        pyrazinyl, each of which may be optionally substituted with a        halo substituent;        (p) Het¹ is a bicyclic heteroaryl selected from        imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl-6-yl or        imidazo[1,2-a]pyridinyl-2-yl;        (q) 7- to 10-membered saturated spirocarbobicyclic system is in        particular

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein R¹ is CF₃.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein R¹ is CF₃, and wherein R² is hydrogen.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein Ar is phenyl optionally substituted according toany of the other embodiments.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein L² is linked to a carbon atom of L¹.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (b).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (c).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (d).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (e).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (f).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (g).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); (b), (d), (e) or (f); and R^(4a) isother than hydrogen. In an embodiment, the present invention relates tothose compounds of Formula (I) and the pharmaceutically acceptablesalts, and the solvates thereof, or any subgroup thereof as mentioned inany of the other embodiments, wherein --L²-R³ is (a) or (f); and R^(4a)is other than hydrogen. In an embodiment, the present invention relatesto those compounds of Formula (I) and the pharmaceutically acceptablesalts, and the solvates thereof, or any subgroup thereof as mentioned inany of the other embodiments, wherein --L²-R³ is (a); and R^(4a) isother than hydrogen.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); and when L² is linked to a nitrogenatom of L¹ then R^(4a) is other than hydrogen.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a) or (f); and when L² is linked to anitrogen atom of L¹ then R⁴ is other than hydrogen.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a), (b), (d), (e) or (f); and when L²is linked to a nitrogen atom of L¹ then R^(4a) is other than hydrogen.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments wherein L¹ represents

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); R³ is Het¹ or Het².

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); R³ is Het¹.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); R³ is Het¹; and Het¹ is azetidinyloptionally substituted as defined in any of the other embodiments.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein --L²-R³ is (a); R³ is Het¹ or Het²; Het¹ is amonocyclic heteroaryl selected from the group consisting of pyridyl, 4-,5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, andimidazolyl; each of which may be optionally substituted with one, two,or three substituents each independently selected from the groupconsisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR²⁷R²⁷, and —C(═O)NR²⁷R^(27b); andHet² is a non-aromatic heterocyclyl selected from the group consistingof azetidinyl, pyrrolidinyl, and piperidinyl; each of which may beoptionally substituted with one, two, or three substituents eachindependently selected from the group consisting of halo, —CN, —OR²⁴,—NR^(27a)R^(27b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein Ar is phenyl which may be optionally substitutedwith one, two, or three substituents each independently selected fromthe group consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), andC₁₋₄alkyl optionally substituted with a substituent selected from thegroup consisting of fluoro, —CN, —OR⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b);

Het¹ is a monocyclic heteroaryl selected from the group consisting ofpyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl,thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl,isothiazolyl, thiadiazolyl, and isoxazolyl; each of which may beoptionally substituted with one, two, or three substituents eachindependently selected from the group consisting of halo, —CN, —OR²⁴,—NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); andHet² is a monocyclic non-aromatic heterocyclyl optionally substitutedwith one, two, or three substituents each independently selected fromthe group consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), andC₁₋₄alkyl optionally substituted with a substituent selected from thegroup consisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b).

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is selected from the group consisting of >CR^(4a)R^(4b), and—CHR^(4a)CHR⁵—; whereinL² is linked to a nitrogen atom of L¹;R^(4a) is selected from the group consisting of —C(═O)NR^(7a)R^(7b); andC-linked 4- to 7-membered non-aromatic heterocyclyl containing at leastone nitrogen, oxygen or sulfur atom;R⁵ is selected from the group consisting of hydrogen; —OR⁶;—NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of fluoro, —CN,—OR⁸, and—NR^(9a)R^(9b);R^(4b) is selected from the group consisting of hydrogen and methyl; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is selected from the group consisting of —C(═O)NR^(7a)R^(7b); andC-linked 4- to 7-membered non-aromatic heterocyclyl containing at leastone nitrogen, oxygen or sulfur atom;R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is selected from the group consisting of —C(═O)NR^(7a)R^(7b); andC-linked 4- to 7-membered non-aromatic heterocyclyl containing at leastone nitrogen, oxygen or sulfur atom;R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, and Het².

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is selected from the group consisting of >CR^(4a)R^(4b), and—CHR^(4a)CHR⁵—; whereinL² is linked to a nitrogen atom of L¹;R^(4a) is —C(═O)NR^(7a)R^(7b);R⁵ is selected from the group consisting of hydrogen; —OR⁶;—NR^(7a)R^(7b);—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b);R^(4a) is selected from the group consisting of hydrogen and methyl; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is —C(═O)NR^(7a)R^(7b);R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is —C(═O)NR^(7a)R^(7b);R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, and Het².

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is selected from the group consisting of >CR^(4a)R^(4b), and—CHR^(4a)CHR⁵—; whereinL² is linked to a nitrogen atom of L¹;R^(4a) is C-linked 4- to 7-membered non-aromatic heterocyclyl containingat least one nitrogen, oxygen or sulfur atom;R⁵ is selected from the group consisting of hydrogen; —OR⁶;—NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of fluoro, —CN,—OR⁸, and—NR^(9a)R^(9b);R^(4b) is selected from the group consisting of hydrogen and methyl; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is C-linked 4- to 7-membered non-aromatic heterocyclyl containingat least one nitrogen, oxygen or sulfur atom;R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, Het¹, Het², and a 7- to10-membered saturated spirocarbobicyclic system.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is (a), whereinL² is >CR^(4a)R^(4b); whereinL² is linked to a nitrogen atom of L¹;R^(4a) is C-linked 4- to 7-membered non-aromatic heterocyclyl containingat least one nitrogen, oxygen or sulfur atom;R^(4b) is hydrogen; andR³ is selected from the group consisting of Ar, and Het².

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein L² is linked to a nitrogen atom of L¹.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is selected from the group consisting of

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

--L²-R³ is selected from the group consisting of

wherein R¹⁸ is hydrogen or methyl.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

L¹ represents a N-linked 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms selected from the group consistingof (a), (b), (c), (d), (e), (f), (g), (h), and (i)

wherein a represents the position of linkage to the thienopyrimidinylheterocycle.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein Het² is monocyclic heterocyclyl optionallysubstituted with one, two, or three substituents as described in theother embodiments.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein Het² is a non-aromatic heterocyclyl selected fromazetidinyl, oxetanyl, tetrahydrofuranyl, piperidinyl, tetrahydropyranyl,tetrahydro-2H-thiopyranyl 1,1-dioxide,

each of which are optionally substituted with one, two, or threesubstituents as described in the other embodiments.

In an embodiment, the present invention relates to those compounds ofFormula (I) and the pharmaceutically acceptable salts, and the solvatesthereof, or any subgroup thereof as mentioned in any of the otherembodiments, wherein

Het² is a non-aromatic heterocyclyl selected from

each of which are optionally substituted with one, two, or threesubstituents as described in the other embodiments.

Particular compounds of Formula (I) are compounds 82, 84, 273, and 274,including the stereoisomeric forms, the pharmaceutically acceptablesalts thereof, in particular the hydrochloride salts thereof, and thesolvates thereof.

Particular compounds of Formula (I) are compounds 82, 84, 273, and 274.

In an embodiment the compound of Formula (I) is selected from the groupconsisting of any of the exemplified compounds,

and the free bases, the pharmaceutically acceptable addition salts, andthe solvates thereof.

All possible combinations of the above-indicated embodiments areconsidered to be embraced within the scope of this invention.

Methods for the Preparation of Compounds of Formula (I)

In this section, as in all other sections unless the context indicatesotherwise, references to Formula (I) also include all other sub-groupsand examples thereof as defined herein.

The general preparation of some typical examples of the compounds ofFormula (I) is described hereunder and in the specific examples, and aregenerally prepared from starting materials which are either commerciallyavailable or prepared by standard synthetic processes commonly used bythose skilled in the art. The following schemes are only meant torepresent examples of the invention and are in no way meant to be alimit of the invention.

Alternatively, compounds of the present invention may also be preparedby analogous reaction protocols as described in the general schemesbelow, combined with standard synthetic processes commonly used by thoseskilled in the art of organic chemistry.

The skilled person will realize that in the reactions described in theSchemes, although this is not always explicitly shown, it may benecessary to protect reactive functional groups (for example hydroxy,amino, or carboxy groups) where these are desired in the final product,to avoid their unwanted participation in the reactions. For example inScheme 1, the NH moiety on the L¹ N-linked 7- to 10-membered saturatedspiroheterobicyclic system containing one or two N-atoms can beprotected with a tert-butoxycarbonyl protecting group. In general,conventional protecting groups can be used in accordance with standardpractice. The protecting groups may be removed at a convenientsubsequent stage using methods known from the art. This is illustratedin the specific examples.

The skilled person will realize that in the reactions described in theSchemes, it may be advisable or necessary to perform the reaction underan inert atmosphere, such as for example under N₂-gas atmosphere.

It will be apparent for the skilled person that it may be necessary tocool the reaction mixture before reaction work-up (refers to the seriesof manipulations required to isolate and purify the product(s) of achemical reaction such as for example quenching, column chromatography,extraction).

The skilled person will realize that heating the reaction mixture understirring may enhance the reaction outcome. In some reactions microwaveheating may be used instead of conventional heating to shorten theoverall reaction time.

The skilled person will realize that another sequence of the chemicalreactions shown in the Schemes below, may also result in the desiredcompound of Formula (I).

The skilled person will realize that intermediates and final compoundsshown in the Schemes below may be further functionalized according tomethods well-known by the person skilled in the art. The intermediatesand compounds described herein can be isolated in free form or as asalt.

Scheme 1

In general, compounds of Formula (I) wherein all variables are definedaccording to the scope of the present invention, can be preparedaccording to the following reaction Scheme 1. In Scheme 1,

represents L¹ as a 7- to 10-membered saturated spiroheterobicyclicsystem containing two N-atoms and which is N-linked to thethienopyrimidinyl heterocycle, LG¹ and LG² each represent a suitableleaving group, such as for example halo or methanesulfonyl; PG¹represents a suitable protecting group, such as for exampletert-butyloxycarbonyl; R^(3a)-PG² represents an R³ as defined in Formula(I) with an appropriate protecting group, such as for exampletert-butyloxycarbonyl, when the R³ substituent bears an amino group. TheX in formula (XI) represents CH or N (in formula (XI) L² can be linkedto a carbon or a N-atom). All other variables in Scheme 1 are definedaccording to the scope of the present invention.

In Scheme 1, the following reaction conditions apply:

1: at a suitable temperature such as ranged from rt to 90° C., in thepresence of a suitable base such as for example diisopropylethylamine,in a suitable solvent such as for example acetonitrile or isopropanol orethanol;2: at a suitable temperature range such as for example from 0° C. toroom temperature, in the presence of suitable cleavage conditions, suchas for example an acid such as HCl or trifluoroacetic acid in a suitablesolvent such as acetonitrile or dichloromethane when PG¹ istert-butyloxycarbonyl;

Alternatively, at a suitable temperature such as for example roomtemperature in a suitable solvent such as acetic acid

3: at a suitable temperature such as for example room temperature orreflux, in the presence of a suitable base such as for example potassiumcarbonate or 1,8-Diazabicyclo[5.4.0]undec-7-ene, in a suitable solventsuch as for example acetonitrile or DMSO;4: at a suitable temperature such as for example room temperature or 90°C., in the presence of a suitable base such as for example potassiumcarbonate or 1,8-Diazabicyclo[5.4.0]undec-7-ene, in a suitable solventsuch as for example acetonitrile or DMSO;5: at a suitable reaction temperature range such as for example from 0°C. to room temperature, in the presence of suitable cleavage conditions,such as for example an acid such as HCl or trifluoroactic acid in asuitable solvent such as acetonitrile or dichloromethane when PG² istert-butyloxycarbonyl.6: at a suitable temperature such as for example at room temperature,eventually in the presence of a suitable base such as for exampletrimethylamine or a suitable acid such as for example acetic acid, in asuitable solvent such as for example anhydrous dichloromethane,dichloroethane or tetrahydropyrane;7: at a suitable temperature, for example room temperature, in thepresence of a suitable reducing agent, such as for example NaBH(OAc)₃,in a suitable solvent such as dichloromethane, dichloroethane ortetrahydropyran; yielding a compound of Formula (I) wherein L¹ is aN-linked 7- to 10-membered saturated spiroheterobicyclic systemcontaining two N-atoms and L² is CH₂.

Steps 6 and 7 can conveniently be performed as a one-pot procedure.

Alternatively, step 6 and 7 can be performed in the presence of asuitable acid such as for example acetic acid, a suitable catalyst suchas platinum oxide, in a suitable solvent such as for example ethanol ata suitable temperature such as for example 60° C.;

8: at a suitable temperature such as for example at 90° C., in thepresence of a suitable base such as for example diisopropylethylamine,in a suitable solvent such as for example acetonitrile or isopropanol.In step 8, reagent of Formula (XI), X represents CH or N, and L² and R³are as defined according to the scope of the invention. Reagents ofFormula (XI) are either commercially available or can be prepared bymethods known to the skilled person from commercially available startingmaterials, e.g. by appropriate protection/deprotection steps andfunctional group interconversion, from starting materials, such as2-azaspiro[3.3]heptan-6-ol (CAS[1256352-97-2]).

Scheme 2

Intermediates of Formula (II), wherein R² is methyl, can be preparedaccording to the following reaction Scheme 2, wherein LG¹ represents asuitable leaving group, such as for example halo or methanesulfonyl. Allother variables in Scheme 2 are defined according to the scope of thepresent invention.

In Scheme 2, the following reaction conditions apply:

1: at a suitable temperature such as for example at reflux temperature,in the presence of acetic anhydride and a suitable base such as forexample trimethylamine, in a suitable solvent such as for exampletoluene;2: at a suitable temperature such as for example at reflux temperature,in the presence of a suitable base such as potassium hydroxide, in asuitable solvent such as for example ethanol;3: under suitable reaction conditions to form a leaving group, such asfor example, chloro, for example by reaction with phosphoryl trichlorideat a suitable temperature such as 110° C.

Scheme 3

In general, compounds of Formula (I-a) wherein the variables are definedaccording to the scope of the present invention, but wherein L² islimited to L²a (the options that can be obtained by this Scheme), can beprepared according to the following reaction Scheme 3. All othervariables in Scheme 3 are defined according to the scope of the presentinvention or as defined before.

In Scheme 3, the following reaction conditions apply:

1: at a suitable temperature such as for example room temperature at 45°C., in the presence of titanium (IV) ethoxide or titanium (IV)isopropoxide, in a suitable solvent such as for exampletetrahydropyrane, dichloroethane or a mixture of dichlorethane andmethanol;

Alternatively, at a suitable temperature such as for example roomtemperature, with or without a suitable acid such as for exampletrifluoroacetic acid, in a suitable solvent such as for exampletetrahydropyrane;

2: at a suitable temperature such as for example room temperature, inthe presence of a suitable reducting agent such as for example sodiumborohydride, sodium triacetoxyborohydride or sodium cyanoborohydride, ina suitable solvent such as for example tetrahydropyrane, dichloroethaneor a mixture of dichloroethane and methanol;

Steps 1 and 2 can be performed as a one-pot procedure.

scheme 4

In general, compounds of Formula (I-b) wherein R^(4a) is restricted toR^(4a1) being C₁₋₄alkyl or a C-linked 4- to 7-membered non-aromaticheterocyclyl containing at least one nitrogen, oxygen or sulfur atom canbe prepared according to the following reaction Scheme 4. In Scheme 4,halo means chloro, bromo or iodo. All other variables in Scheme 4 aredefined according to the scope of the present invention or as definedbefore.

In Scheme 4, the following reaction conditions apply:

1: at a suitable temperature such as for example room temperature or 45°C., in the presence of titanium (IV) ethoxide or titanium (IV)isopropoxide, in a suitable solvent such as for example tetrahydropyran;2: at a suitable temperature ranged from 0° C. to room temperature, in asuitable solvent such as for example tetrahydrofurane.

Steps 1 and 2 can be performed as a one-pot procedure.

Scheme 5

In general, compounds of Formula (Ic) wherein R³ is restricted to R^(3c)being

can be prepared according to the following reaction Scheme 5. All othervariables in Scheme 5 are defined according to the scope of the presentinvention or as defined before.

In Scheme 5, L² is linked to a N-atom of L¹.

In Scheme 5, the following reaction conditions apply:

1: at a suitable temperature, such as for example room temperature, inthe presence of a suitable acid coupling agent, such as for example1-[bis(dimethylamino)methylene]-1H-benzotriazoliumhexafluorophosphate(1-)3-oxide(HBTU) or1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU), in the presence of a suitable basesuch as for example N-ethyl-N-(1-methylethyl)-2-propanamine (DIPEA), ina suitable solvent such as N,N-dimethylformamide (DMF);

Scheme 6

In general, compounds of Formula (Id) wherein L² is restricted to SO₂,can be prepared according to the following reaction Scheme 6. All othervariables in Scheme 6 are defined according to the scope of the presentinvention or as defined before. In Scheme 6, L² (>SO₂ in Scheme 6) islinked to a N-atom of L¹.

In Scheme 6, the following reaction conditions apply:

1; at a suitable temperature, for example room temperature, in thepresence of a suitable base such as for example potassium carbonate, ina suitable solvent such as for example acetonitrile.

Scheme 7

In general, compounds of Formula (Ie) and (If) can be prepared accordingto the following reaction Scheme 7. Both in (Ie) and (If) the L² part ofthe molecule is linked to a nitrogen atom of L¹. All other variables aredefined according to the scope of the present invention or as definedbefore.

In Scheme 7, the following reaction conditions apply:

1: at a suitable temperature such as for example 60° C., in a suitablesolvent such as for example ethanol.

It will be appreciated that where appropriate functional groups exist,compounds of various formulae or any intermediates used in theirpreparation may be further derivatised by one or more standard syntheticmethods employing condensation, substitution, oxidation, reduction, orcleavage reactions. Particular substitution approaches includeconventional alkylation, arylation, heteroarylation, acylation,sulfonylation, halogenation, nitration, formylation and couplingprocedures.

The compounds of Formula (I) may be synthesized in the form of racemicmixtures of enantiomers which can be separated from one anotherfollowing art-known resolution procedures. The racemic compounds ofFormula (I) containing a basic nitrogen atom may be converted into thecorresponding diastereomeric salt forms by reaction with a suitablechiral acid. Said diastereomeric salt forms are subsequently separated,for example, by selective or fractional crystallization and theenantiomers are liberated therefrom by alkali. An alternative manner ofseparating the enantiomeric forms of the compounds of Formula (I)involves liquid chromatography using a chiral stationary phase. Saidpure stereochemically isomeric forms may also be derived from thecorresponding pure stereochemically isomeric forms of the appropriatestarting materials, provided that the reaction occursstereospecifically.

In the preparation of compounds of the present invention, protection ofremote functionality (e.g., primary or secondary amine) of intermediatesmay be necessary. The need for such protection will vary depending onthe nature of the remote functionality and the conditions of thepreparation methods. Suitable amino-protecting groups (NH-Pg) includeacetyl, trifluoroacetyl, t-butoxycarbonyl (Boc), benzyloxycarbonyl (CBz)and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protectionis readily determined by one skilled in the art. For a generaldescription of protecting groups and their use, see T. W. Greene and P.G. M. Wuts, Protective Groups in Organic Synthesis, 4th ed., Wiley,Hoboken, N.J., 2007.

Pharmacology

It has been found that the compounds of the present invention block theinteraction of menin with MLL proteins and oncogenic MLL fusionproteins. Therefore the compounds according to the present invention andthe pharmaceutical compositions comprising such compounds may be usefulfor the treatment or prevention, in particular treatment, of diseasessuch as cancer, myelodysplastic syndrome (MDS) and diabetes.

In particular, the compounds according to the present invention and thepharmaceutical compositions thereof may be useful in the treatment orprevention of cancer. According to one embodiment, cancers that maybenefit from a treatment with menin/MLL inhibitors of the inventioncomprise leukemias, myeloma or a solid tumor cancer (e.g. prostatecancer, lung cancer, breast cancer, pancreatic cancer, colon cancer,liver cancer, melanoma and glioblastoma, etc.). In some embodiments, theleukemias include acute leukemias, chronic leukemias, myeloid leukemias,myelogeneous leukemias, lymphoblastic leukemias, lymphocytic leukemias,Acute myelogeneous leukemias (AML), Chronic myelogenous leukemias (CML),Acute lymphoblastic leukemias (ALL), Chronic lymphocytic leukemias(CLL), T cell prolymphocytic leukemias (T-PLL), Large granularlymphocytic leukemia, Hairy cell leukemia (HCL), MLL-rearrangedleukemias, MLL-PTD leukemias, MLL amplified leukemias, MLL-positiveleukemias, leukemias exhibiting HOX/MEIS1 gene expression signaturesetc.

Hence, the invention relates to compounds of Formula (I), the tautomersand the stereoisomeric forms thereof, and the pharmaceuticallyacceptable salts, and the solvates thereof, for use as a medicament.

The invention also relates to the use of a compound of Formula (I), atautomer or a stereoisomeric form thereof, or a pharmaceuticallyacceptable salt, or a solvate thereof, or a pharmaceutical compositionaccording to the invention, for the manufacture of a medicament.

The present invention also relates to a compound of Formula (I), atautomer or a stereoisomeric form thereof, or a pharmaceuticallyacceptable salt, or a solvate thereof, or a pharmaceutical compositionaccording to the invention, for use in the treatment, prevention,amelioration, control or reduction of the risk of disorders associatedwith the interaction of menin with MLL proteins and oncogenic MLL fusionproteins in a mammal, including a human, the treatment or prevention ofwhich is affected or facilitated by blocking the interaction of meninwith MLL proteins and oncogenic MLL fusion proteins.

Also, the present invention relates to the use of a compound of Formula(I), a tautomer or a stereoisomeric form thereof, or a pharmaceuticallyacceptable salt, or a solvate thereof, or a pharmaceutical compositionaccording to the invention, for the manufacture of a medicament fortreating, preventing, ameliorating, controlling or reducing the risk ofdisorders associated with the interaction of menin with MLL proteins andoncogenic MLL fusion proteins in a mammal, including a human, thetreatment or prevention of which is affected or facilitated by blockingthe interaction of menin with MLL proteins and oncogenic MLL fusionproteins.

The invention also relates to a compound of Formula (I), a tautomer or astereoisomeric form thereof, or a pharmaceutically acceptable salt, or asolvate thereof, for use in the treatment or prevention of any one ofthe diseases mentioned hereinbefore.

The invention also relates to a compound of Formula (I), a tautomer or astereoisomeric form thereof, or a pharmaceutically acceptable salt, or asolvate thereof, for use in treating or preventing any one of thediseases mentioned hereinbefore.

The invention also relates to the use of a compound of Formula (I), atautomer or a stereoisomeric form thereof, or a pharmaceuticallyacceptable salt, or a solvate thereof, for the manufacture of amedicament for the treatment or prevention of any one of the diseaseconditions mentioned hereinbefore.

The compounds of the present invention can be administered to mammals,preferably humans, for the treatment or prevention of any one of thediseases mentioned hereinbefore.

In view of the utility of the compounds of Formula (I), the tautomersand the stereoisomeric forms thereof, and the pharmaceuticallyacceptable salts, and the solvates thereof, there is provided a methodof treating warm-blooded animals, including humans, suffering from anyone of the diseases mentioned hereinbefore.

Said method comprises the administration, i.e. the systemic or topicaladministration, preferably oral administration, of a therapeuticallyeffective amount of a compound of Formula (I), a tautomer or astereoisomeric form thereof, or a pharmaceutically acceptable salt, or asolvate thereof, to warm-blooded animals, including humans.

Therefore, the invention also relates to a method for the treatment orprevention of any one of the diseases mentioned hereinbefore comprisingadministering a therapeutically effective amount of compound accordingto the invention to a patient in need thereof.

One skilled in the art will recognize that a therapeutically effectiveamount of the compounds of the present invention is the amountsufficient to have therapeutic activity and that this amount variesinter alias, depending on the type of disease, the concentration of thecompound in the therapeutic formulation, and the condition of thepatient. Generally, the amount of a compound of the present invention tobe administered as a therapeutic agent for treating the disordersreferred to herein will be determined on a case by case by an attendingphysician.

Those of skill in the treatment of such diseases could determine theeffective therapeutic daily amount from the test results presentedhereinafter. An effective therapeutic daily amount would be from about0.005 mg/kg to 100 mg/kg, in particular 0.005 mg/kg to 50 mg/kg, inparticular 0.01 mg/kg to 50 mg/kg body weight, more in particular from0.01 mg/kg to 25 mg/kg body weight, preferably from about 0.01 mg/kg toabout 15 mg/kg, more preferably from about 0.01 mg/kg to about 10 mg/kg,even more preferably from about 0.01 mg/kg to about 1 mg/kg, mostpreferably from about 0.05 mg/kg to about 1 mg/kg body weight. Aparticular effective therapeutic daily amount might be 1 mg/kg bodyweight, 2 mg/kg body weight, 4 mg/kg body weight, or 8 mg/kg bodyweight. The amount of a compound according to the present invention,also referred to herein as the active ingredient, which is required toachieve a therapeutically effect may vary on case-by-case basis, forexample with the particular compound, the route of administration, theage and condition of the recipient, and the particular disorder ordisease being treated. A method of treatment may also includeadministering the active ingredient on a regimen of between one and fourintakes per day. In these methods of treatment the compounds accordingto the invention are preferably formulated prior to administration. Asdescribed herein below, suitable pharmaceutical formulations areprepared by known procedures using well known and readily availableingredients.

The present invention also provides compositions for preventing ortreating the disorders referred to herein. Said compositions comprisinga therapeutically effective amount of a compound of Formula (I), atautomer or a stereoisomeric form thereof, or a pharmaceuticallyacceptable salt, or a solvate thereof, and a pharmaceutically acceptablecarrier or diluent.

While it is possible for the active ingredient to be administered alone,it is preferable to present it as a pharmaceutical composition.Accordingly, the present invention further provides a pharmaceuticalcomposition comprising a compound according to the present invention,together with a pharmaceutically acceptable carrier or diluent. Thecarrier or diluent must be “acceptable” in the sense of being compatiblewith the other ingredients of the composition and not deleterious to therecipients thereof.

The pharmaceutical compositions of this invention may be prepared by anymethods well known in the art of pharmacy, for example, using methodssuch as those described in Gennaro et al. Remington's PharmaceuticalSciences (18^(th) ed., Mack Publishing Company, 1990, see especiallyPart 8: Pharmaceutical preparations and their Manufacture). Atherapeutically effective amount of the particular compound, in baseform or salt form, as the active ingredient is combined in intimateadmixture with a pharmaceutically acceptable carrier, which may take awide variety of forms depending on the form of preparation desired foradministration. These pharmaceutical compositions are desirably inunitary dosage form suitable, preferably, for systemic administrationsuch as oral, percutaneous or parenteral administration; or topicaladministration such as via inhalation, a nose spray, eye drops or via acream, gel, shampoo or the like. For example, in preparing thecompositions in oral dosage form, any of the usual pharmaceutical mediamay be employed, such as, for example, water, glycols, oils, alcoholsand the like in the case of oral liquid preparations such assuspensions, syrups, elixirs and solutions: or solid carriers such asstarches, sugars, kaolin, lubricants, binders, disintegrating agents andthe like in the case of powders, pills, capsules and tablets. Because oftheir ease in administration, tablets and capsules represent the mostadvantageous oral dosage unit form, in which case solid pharmaceuticalcarriers are obviously employed. For parenteral compositions, thecarrier will usually comprise sterile water, at least in large part,though other ingredients, for example, to aid solubility, may beincluded. Injectable solutions, for example, may be prepared in whichthe carrier comprises saline solution, glucose solution or a mixture ofsaline and glucose solution. Injectable suspensions may also be preparedin which case appropriate liquid carriers, suspending agents and thelike may be employed. In the compositions suitable for percutaneousadministration, the carrier optionally comprises a penetration enhancingagent and/or a suitable wettable agent, optionally combined withsuitable additives of any nature in minor proportions, which additivesdo not cause any significant deleterious effects on the skin. Saidadditives may facilitate the administration to the skin and/or may behelpful for preparing the desired compositions. These compositions maybe administered in various ways, e.g., as a transdermal patch, as aspot-on or as an ointment.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient calculated to produce the desiredtherapeutic effect in association with the required pharmaceuticalcarrier. Examples of such dosage unit forms are tablets (includingscored or coated tablets), capsules, pills, powder packets, wafers,injectable solutions or suspensions, teaspoonfuls, tablespoonfuls andthe like, and segregated multiples thereof.

The present compounds can be used for systemic administration such asoral, percutaneous or parenteral administration; or topicaladministration such as via inhalation, a nose spray, eye drops or via acream, gel, shampoo or the like. The compounds are preferably orallyadministered. The exact dosage and frequency of administration dependson the particular compound of Formula (I) used, the particular conditionbeing treated, the severity of the condition being treated, the age,weight, sex, extent of disorder and general physical condition of theparticular patient as well as other medication the individual may betaking, as is well known to those skilled in the art. Furthermore, it isevident that said effective daily amount may be lowered or increaseddepending on the response of the treated subject and/or depending on theevaluation of the physician prescribing the compounds of the instantinvention.

The compounds of the present invention may be administered alone or incombination with one or more additional therapeutic agents. Combinationtherapy includes administration of a single pharmaceutical dosageformulation which contains a compound according to the present inventionand one or more additional therapeutic agents, as well as administrationof the compound according to the present invention and each additionaltherapeutic agent in its own separate pharmaceutical dosage formulation.For example, a compound according to the present invention and atherapeutic agent may be administered to the patient together in asingle oral dosage composition such as a tablet or capsule, or eachagent may be administered in separate oral dosage formulations.

Therefore, an embodiment of the present invention relates to a productcontaining as first active ingredient a compound according to theinvention and as further active ingredient one or more anticancer agent,as a combined preparation for simultaneous, separate or sequential usein the treatment of patients suffering from cancer.

The one or more other medicinal agents and the compound according to thepresent invention may be administered simultaneously (e.g. in separateor unitary compositions) or sequentially in either order. In the lattercase, the two or more compounds will be administered within a period andin an amount and manner that is sufficient to ensure that anadvantageous or synergistic effect is achieved. It will be appreciatedthat the preferred method and order of administration and the respectivedosage amounts and regimes for each component of the combination willdepend on the particular other medicinal agent and compound of thepresent invention being administered, their route of administration, theparticular condition, in particular tumour being treated and theparticular host being treated. The optimum method and order ofadministration and the dosage amounts and regime can be readilydetermined by those skilled in the art using conventional methods and inview of the information set out herein.

The weight ratio of the compound according to the present invention andthe one or more other anticancer agent(s) when given as a combinationmay be determined by the person skilled in the art. Said ratio and theexact dosage and frequency of administration depends on the particularcompound according to the invention and the other anticancer agent(s)used, the particular condition being treated, the severity of thecondition being treated, the age, weight, gender, diet, time ofadministration and general physical condition of the particular patient,the mode of administration as well as other medication the individualmay be taking, as is well known to those skilled in the art.Furthermore, it is evident that the effective daily amount may belowered or increased depending on the response of the treated subjectand/or depending on the evaluation of the physician prescribing thecompounds of the instant invention. A particular weight ratio for thepresent compound of Formula (I) and another anticancer agent may rangefrom 1/10 to 10/1, more in particular from 1/5 to 5/1, even more inparticular from 1/3 to 3/1.

The following examples further illustrate the present invention.

EXAMPLES

Several methods for preparing the compounds of this invention areillustrated in the following examples. Unless otherwise noted, allstarting materials were obtained from commercial suppliers and usedwithout further purification.

Hereinafter, the terms: ‘ACN or ‘MeCN’ means acetonitrile, ‘DCM’ meansdichloromethane, ‘DIPEA’ means N,N-diisopropylethylamine, ‘DIPE or‘DiPE” means diisopropyl ether, ‘h’ means hours(s), ‘min’ meansminute(s), ‘DMF’ means dimethylformamide, ‘DSC’ means differentialscanning calorimetry, ‘TEA or ‘Et₃N’ means triethyl amine, ‘Et₂O’ meansdiethyl ether, ‘EtOAc’ or ‘EA’ means ethyl acetate, ‘EtOH’ meansethanol, ‘HPLC’ means High-performance Liquid Chromatography, ‘iPrOH’means isopropyl alcohol, ‘LC/MS’ means Liquid Chromatography/MassSpectrometry, ‘MeOH’ means methanol, ‘NMR’ means Nuclear MagneticResonance, ‘rt or ‘RT’ means room temperature, ‘SFC’ means supercriticalfluid chromatography, ‘OR’ means optical rotation, ‘sat. aq.’ meanssaturated aqueous. ‘AcCl’ means acetyl chloride, ‘AcOH’ or ‘HOAc’ meansacetic acid, ‘BOC’ or ‘Boc’ means tert-butyloxycarbonyl, ‘Celite®’ meansdiatomaceous earth, ‘CH₃COONH₄’ means ammonium acetate, ‘COMU®’ means(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate, ‘CO₂’ means carbon dioxide, ‘DCE’ meansdichloroethane, ‘DMAP’ means dimethylaminopyridine, ‘DMSO’ meansdimethyl sulfoxyde, ‘DBU’ means 1,8-diazabicyclo[5.4.0]undecene-7,‘EDCI.HCl’ means 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, ‘ee’ means enantiomeric excess, ‘eq.’ or ‘equiv.’ meansequivalent(s), ‘EtMgBr’ means ethyl magnesium bromide, ‘Et₂O’ meansdiethyl ether, ‘EtOAc’ means ethyl acetate, ‘Et₃N’ or ‘TEA’ meanstriethylamine, ‘EtOH’ means ethanol, ‘h’ means hours(s), ‘HATU’ meansO-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluroniumhexafluorophosphate, ‘HCl’ means hydrochloric acid, ‘HOBT’ meansN-Hydroxybenzotrizole monohydrate, ‘H₂O’ means water, ‘iPrMgCl’ meansisopropyl magnesium chloride, ‘iPrNH₂’ means isopropylamine, ‘K₂CO₃’means potassium carbonate, ‘Me-THF’ means 2-methyl-tetrahydrofuran,‘MeMgBr’ or ‘CH₃MgBr’ means methyl magnesium bromide, ‘MeOH’ meansmethanol, ‘MgSO₄’ means magnesium sulfate, ‘min’ means minute(s),‘NaBH(OAc)₃’ means sodium triacetoxyborohydride, ‘NaBH₃CN’ means sodiumcyanoborohydride, ‘Na₂CO₃’ means sodium carbonate, ‘NaH’ means sodiumhydride, ‘NaHCO₃’ means sodium hydrogenocarbonate, ‘NaOH’ meanspotassium hydroxide, ‘Na₂SO₄’ means sodium sulfate, ‘NH₄Cl’ meansammonium chloride, ‘NH₄HCO₃’ means ammonium bicarbonate, ‘NH₄OH’ meansammonia solution 30% aqueous, ‘Quant. or quant’ means quantitative,‘R_(t)’ means retention time, ‘SFC’ means supercritical fluidchromatography, ‘T’ means temperature, ‘TBAF’ means tetrabutylammoniumfluoride, ‘TBDMS’ or ‘SMDBT’ means tert-butyldimethylsilyl, ‘TFA’ or‘CF₃COOH’ means trifluoroacetic acid, ‘THF’ means tetrahydrofuran,‘Ti(OEt)₄’ means titanium ethoxyde, ‘Ti(OiPr)₄’ means titaniumisopropoxide, ‘v.’ means volume, ‘F₃C’ or ‘CF₃’ means trifluoromethyl,‘HBTU’ means1-[bis(dimethylamino)methylene]-1H-benzotriazoliumhexafluorophosphate(1-)3-oxide.

As understood by a person skilled in the art, compounds synthesisedusing the protocols as indicated may exist as a solvate e.g. hydrate,and/or contain residual solvent or minor impurities. Compounds isolatedas a salt form, may be integer stoichiometric i.e. mono- or di-salts, orof intermediate stoichiometry.

The stereochemical configuration for centres in some compounds may bedesignated “R” or “S” when the mixture(s) was separated; for somecompounds, the stereochemical configuration at indicated centres hasbeen designated as “*R” (first eluted from the column in case the columnconditions are described in the synthesis protocol and when only onestereocentre present) or “*S” (second eluted from the column in case thecolumn conditions are described in the synthesis protocol and when onlyone stereocentre present) when the absolute stereochemistry isundetermined (even if the bonds are drawn stereospecifically) althoughthe compound itself has been isolated as a single stereoisomer and isenantiomerically pure.

For example, it will be clear that compound 179

is

Compounds having two stereocentres of which only the stereochemicalconfiguration of one stereocentre is indicated by * (e.g. *R or *S) (seefor example compound 186 or 281), follow a similar rule as above. Thismeans that the absolute stereoconfiguration of the stereocentreindicated by * is undetermined (even if the bonds are drawnstereospecifically) although the compound is enantiomerically pure atthe indicated centre.

For compounds such as 188, 189, 190, 191, 235, 236, 237, and 238,wherein the stereochemical configuration of two stereocentres isindicated by * (e.g. *R or *S), the absolute stereochemistry of thestereocentres is undetermined (even if the bonds are drawnstereospecifically), although the compound itself has been isolated as asingle stereoisomer and is enantiomerically pure. In this case, theconfiguration of the first stereocentre is independent of theconfiguration of the second stereocentre in the same compound.

For example, for Compound 188

this means that the compound is

The paragraphs above about stereochemical configurations, also apply tointermediates.

The term “enantiomerically pure” as used herein means that the productcontains at least 80% by weight of one enantiomer and 20% by weight orless of the other enantiomer. Preferably the product contains at least90% by weight of one enantiomer and 10% by weight or less of the otherenantiomer. In the most preferred embodiment the term “enantiomericallypure” means that the composition contains at least 99% by weight of oneenantiomer and 1% or less of the other enantiomer.

When an intermediate or compound in the experimental part below isindicated as ‘HCl salt’, ‘HCOOH salt’ or ‘TFA salt’ without indicationof the number of equivalents of HCl or TFA, this means that the numberof equivalents of HCl or TFA was not determined.

A skilled person will realize that, even where not mentioned explicitlyin the experimental protocols below, typically after a columnchromatography purification, the desired fractions were collected andthe solvent was evaporated.

In case no stereochemistry is indicated in the spirocycle represented byL1, this means it is a mixture of stereoisomers, unless otherwise isindicated or is clear from the context.

When a stereocentre is indicated with ‘RS’ this means that a racemicmixture was obtained at the indicated centre, unless otherwiseindicated.

A. Preparation of the Intermediates Preparation of Intermediate 1

A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(525 mg, 2.08 mmol) prepared as described in Journal of MedicinalChemistry (2016), 59(3), 892-913, tert-butyl2,7-diazaspiro[4.5]decane-2-carboxylate (550 mg, 2.29 mmol) and DIPEA(1.43 mL, 8.3 mmol) in ACN (12 mL) was heated at 80° C. overnight. Thesolution was cooled and the mixture was poured into cooled water, theproduct was extracted with EtOAc, the organic layer was dried overMgSO₄, filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (stationary phase: irregular 15-40 μm 50g, mobile phase: DCM/MeOH: gradient from 100/0 to 99/1). The productcontaining fractions were collected and evaporated to dryness yielding770 mg (yield 81%) of intermediate 1.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 1, starting fromthe respective starting materials

Intermediate number Structure Quantity (mg) Yield (%) Intermediate 2(from CAS[336191-17-4] and [1628317-85-0])

350 100 Intermediate 3 (from CAS[885270-84-8] and [1628317-85-0])

200 73 Intermediate 4 (from CAS[885270-86-0] and [1628317-85-0])

660 78 Intermediate 5 (from CAS[885268-42-8] and intermediate 15)

355 57

Preparation of Intermediate 3

A solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(11.4 g; 44.96 mmol), ter butyl 2,6 diazaspiro[3.4]octane-2-carboxylate(10.5 g; 49.46 mmol) and DIPEA (15.5 mL; 89.93 mmol) in iPrOH (183 mL)was heated at 90° C. overnight. The solution was cooled to rt and thesolution was poured into water then extracted with EtOAc (3×). Theorganic layer was washed with brine, dried over MgSO₄ and filtered off.

A precipitate (in aqueous layer) was filtered off, washed with few DCMand combined with a previous filtrate. The solvent was evaporated togive 19.9 g of brown solid. The residue was taken up with diethylether,the precipitate was filtered and dried to give 18.5 g of pale brownsolid of intermediate 3 (96%).

Alternative Preparation of Intermediate 3

To a mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(3.00 g, 11.9 mmol) and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (2.5 g, 11.8 mmol) in EtOH (50 mL) was added DIPEA(2 g, 15.5 mmol) in one portion. The mixture was stirred at roomtemperature for 18 h. The mixture was evaporated and the residue wasdiluted in EA (200 mL). The solution was washed with water (100 mL*2),dried over Na₂SO₄, filtered and evaporated to give intermediate 3 (5.10g, 11.9 mmol, 100% yield) as brown oil.

Preparation of Intermediate 6

A mixture of intermediate 1 (770 mg, 1.69 mmol), and a solution of 4NHCl in dioxane (4.22 mL, 16.9 mmol) in ACN (45 mL) was stirred at rtovernight. The mixture was poured out into iced water, basified with 3NNaOH, the product was extracted with DCM, the organic layer was driedover MgSO₄, evaporated to dryness providing 670 mg of intermediate 6which was used without further purification for the next step.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 6, starting fromthe respective starting materials.

Quantity Intermediate number Structure (mg) Yield (%) Intermediate 7(from intermediate 2)

320 Intermediate 8 (from intermediate 4)

582

Preparation of Intermediate 9

In a sealed tube,4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (0.6 g, 2.37mmol) prepared as described in Journal of Medicinal Chemistry (2016),59(3), 892-913, tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate(0.57 g, 2.85 mmol), DIPEA (0.82 mL, 4.75 mmol) in iPrOH(15 mL) wereheated at 90° C. for 2 h. The solution was cooled to rt and the reactionmixture was poured into water then extracted with EtOAc. The organiclayer was washed with water, dried over MgSO₄, filtered and evaporatedto dryness. The crude product was crystallized from Et₂O providing 0.6 g(yield 61%) of intermediate 9.

Preparation of Intermediate 10

A mixture of intermediate 9 (4.43 g; 10.69 mmol) in formic acid (24 mL)was stirred at RT overnight. The reaction mixture was evaporated. Theresidue was taken up twice with Et₂O and evaporated to dryness. Theresidue was purified by chromatography over silica gel (irregular SiOH;80 g; mobile phase: 90% DCM, 10% MeOH, 1% NH₄OH). The pure fractionswere collected and evaporated to dryness yielding 3.34 g (99%) ofintermediate 10.

Preparation of Intermediate 10b

A mixture of intermediate 9 (0.55 g, 1.33 mmol) in formic acid (3 mL)was stirred at RT for 20 h. The mixture was evaporated in vacuo to givea residue that was taken-up twice with Et₂O and evaporated to drynessgiving 0.4 g (yield 96%) of intermediate 10b (formic acid salt). Thecrude product was used without any further purification in the nextstep.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 10b, startingfrom the respective starting materials.

Intermediate number Structure Intermediate 12 (from intermediate 5)

Preparation of Intermediate 11

A mixture of intermediate 3 (8.57 g; 20 mmol) in formic acid (51 mL) wasstirred at rt for 20 h. The reaction mixture was stirred at rt for theweek-end. The mixture was evaporated and the residue was cooled to 5°C., taken-up with DCM and neutralized with aqueous solution of NaOH 3N.The organic layer was washed with water, dried over MgSO₄, filtered andthe solvent was evaporated. The residue (7.63 g of orange oil) waspurified by chromatography over silica gel (irregular bare silica 120 g,mobile phase: 1% NH₄OH, 85% DCM, 15% MeOH). The pure fractions werecollected and the solvent was evaporated to give 3.65 g of yellow oilintermediate 11 (56%).

Alternative Preparation of Intermediate 11

TFA (17.9 mL; 233.38 mmol) was added to a solution of intermediate 3 (5g; 11.67 mmol) in DCM (130 mL) at 5° C. and the reaction mixture wasstirred at rt for 4 h. The reaction mixture was diluted with heptane andevaporated to dryness (3×) to give 10.7 g of brown oil. The residue waspurified by chromatography over silica gel (irregular SiOH 40 μm; 220 g,mobile phase: 1% NH₄OH, 90% DCM, 10% MeOH). The pure fractions werecollected and the solvent was evaporated. The residue (4 g) wassolubilized with DCM and the product was crystallized. The mixture wasevaporated and taken up several times with ACN and the solvent wasevaporated to give 4 g of pale yellow solid intermediate 11.

Preparation of Intermediate 11 b

TFA (2.2 mL; 28 mmol) was added to a solution of intermediate 3 (600 mg;1.4 mmol) in DCM (13 mL) at 0° C. then the reaction mixture was stirredat rt overnight. The reaction mixture was evaporated till dryness togive 1.26 g of intermediate 11b as TFA salt. The product was used itdirectly without purification.

Preparation of Intermediate 11c

A solution of HCl 4M in dioxane (150 mL) was added to intermediate 3(6.5 g; 15.17 mmol) at rt. The reaction mixture was stirred at rt for 1h. The mixture was evaporated in vacuum to give 5.7 g of yellow solidintermediate 11c as HCl salt. The product was used without purificationfor the next step.

Preparation of Intermediate 13

Acetic anhydride (1 mL, 10.7 mmol) was added dropwise at rt, to asolution of 2-amino-5-(2,2,2-trifluoroethyl)thiophene-3-carboxamide (2g, 8.92 mmol) in toluene (50 mL) and trimethylamine (6.2 mL, 44.6 mmol).The solution was heated at reflux for 5 h, poured into water, extractedwith EtOAc, and washed with brine (×2). The organic layer was dried overMgSO₄, filtered and evaporated to dryness, the crude product wastaken-up with Et₂O and the precipitate was filtered to provide 1.5 g ofintermediate 13 (yield 63%/brown solid).

Preparation of Intermediate 14

To a solution of intermediate 13 (1.5 g, 5.63 mmol) in EtOH (70 mL) atrt, was added dropwise a 1M solution of KOH. The reaction mixture wasstirred at rt for 30 min, then the mixture was heated at reflux for 3 h.The reaction mixture was cooled to rt then poured into ice water,acidified with 3N HCl, extracted with DCM and decanted. The combinedorganic layers were washed with brine and dried over MgSO₄, filtered andevaporated to dryness. The residue was crystallized from Et₂O to give0.7 g of intermediate 14 (yield 50%) that was used without furtherpurification in the next step.

Preparation of Intermediate 15

Intermediate 14 (0.7 g, 2.82 mmol) and phosphoryl trichloride (5 mL)were heated at 110° C. for 2 h. The reaction mixture was cooled to rt,then evaporated to dryness. The residue was taken-up carefully with iceand DCM, basified with an aqueous solution of K₂CO₃ (10%) and theorganic layer was washed with water, dried over MgSO₄, filtered andevaporated to dryness to give 0.75 g (yield 99%) of intermediate 15,that was used without further purification in the next step.

Preparation of Intermediate 16 Spiro[3.3]heptan-2-ylmethylmethanesulfonate

To a solution of spiro[3.3]heptan-2-ylmethanol (153 mg, 1.08 mmol) in 4mL of DCM was added TEA (0.464 mL, 3.2 mmol) and the reaction mixturewas cooled to 0° C. Methylsulfonylchloride (0.184 g, 1.605 mmol) wasthen added dropwise, the mixture was allowed to warm to rt and stirredfor 2 h. An aqueous solution of saturated NaHCO₃ (30 mL) and DCM (30 mL)were added. The mixture was separated, the organic layer was collected,washed with brine (10 mL), dried over Na₂SO₄, and evaporated to give 300mg of intermediate 16 as a yellow oil which was used without furtherpurification in the next step.

Preparation of Intermediate 17

To a solution of intermediate 11c (400 mg) and TEA (0.38 mL, 2.76 mmol)under a N₂ flow in DCM (20 mL) was added1-[2-(acetyloxy)ethyl]-1H-pyrrole-2-carboxaldehyde (200 mg, 1.11 mmol).The mixture was stirred at rt for 4 h. NaBH(OAc)₃ (390 mg, 1.84 mmol)was added and the mixture was stirred at rt for 48 h. Then, it waspoured into ice water and the mixture was separated and the aqueouslayer was extracted with DCM. The organic layers were combined, washedwith brine then dried over MgSO₄ and evaporated to dryness. The residuewas purified by chromatography over silica gel (stationary phase:irregular SiOH 15-40 μm 24 g, mobile phase: DCM/MeOH: 97/3). The purefractions were collected and the solvent was evaporated under vacuumyielding 180 mg of intermediate 17.

Preparation of Intermediate 35

1H-pyrazole-4-carbaldehyde (0.5 g; 5.2 mmol) and cesium carbonate (3.39g; 10.4 mmol) were diluted in ACN (10 mL). Then, 2-bromoethyl methylether (0.636 mL; 6.77 mmol) was added and the reaction mixture wasrefluxed for 2 hours. The reaction mixture was partitioned between asaturated solution of NaHCO₃ and EtOAc. The organic layer was separated,dried over MgSO₄, filtered and concentrated.

The residue was purified by silica gel chromatography (irregular SiO₂,120 g, DCM/MeOH: 100/0 to 95/5). The fractions containing the productwere mixed and concentrated to afford 439 mg (55%) of intermediate 35.

Preparation of Intermediate 20

Intermediate 11 (150 mg, 0.46 mmol), (+/−)-methylalpha-bromophenylacetate (0.08 mL, 0.50 mmol) and K₂CO₃ (127 mg; 0.92mmol) in DMF (10 mL) were stirred at rt for 5 h. The reaction mixturewas poured into ice water and EtOAc was added. The organic layer wasseparated, washed with brine, dried over MgSO₄, filtered and evaporatedtill dryness. The residue was purified by chromatography over silica gel(stationary phase: irregular SiOH 15-40 μm 24 g, mobile phase: DCM/MeOH(+10% NH₄OH): gradient from 97/3 to 95/5). The pure fractions werecollected and evaporated to dryness yielding 162 mg (yield 74%) ofintermediate 20.

Preparation of Intermediate 32

Lithium hydroxide monohydrate (71 mg; 1.7 mmol) was added, at rt, to asolution of intermediate 20 (162 mg; 0.34 mmol) in THF (3 mL) and water(3 mL). The mixture was stirred at rt overnight, then concentrated andacidified with an aqueous solution of HCl 3N (pH=2-4).The precipitatewas filtered and dry to give 33 mg (21%) of intermediate 32 (90% ofpurity based on LC/MS). The mother layer was evaporated till dryness togive 243 mg of an impure fraction of intermediate 32.

Preparation of Intermediate 54

Intermediate 11c (333 mg), methyl 2-formylbenzoate (148.5 mg; 0.905mmol), NaBH(OAc)₃ (872 mg; 4.11 mmol) and trimethylamine (250 mg; 2.47mmol) were mixed in dichloroethane (16 mL) and the reaction was stirredat RT overnight. Then, an aqueous solution of NaHCO₃ (1 (mL) was addedand the mixture was extracted with DCM (4*15 mL). The organic layerswere separated, mixed, dried over MgSO₄, filtered and concentrated toafford 450 mg of intermediate 54 as white solid.

The intermediates in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 54, startingfrom the respective starting materials.

Quantity Yield Intermediate number Structure (mg) (%) Intermediate 44(from intermediate 11c and intermediate 52)

340

Preparation of Intermediate 47

A mixture of intermediate 10b (150 mg),1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carboxaldehyde (225 mg; 1.249mmol) and AcOH (24 μL; 0.416 mmol) in dichloroethane (4.5 mL) wasstirred at 50° C. for 2 hours. The reaction mixture was cooled to roomtemperature and NaBH(OAc)₃ (265 mg; 1.249 mmol) was added. The reactionmixture was stirred at room temperature overnight, poured onto a 10%aqueous solution of K₂CO₃ and extracted with DCM. The organic layer wasdecanted, dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (irregular SiOH,24 g; mobile phase: gradient from 0% MeOH, 100% DCM to 10% MeOH, 90%DCM). The pure fractions were collected and evaporated to drynessyielding 150 mg of intermediate 47.

The intermediates in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 47, startingfrom the respective starting materials.

Quantity Yield Intermediate number Structure (mg) (%) Intermediate 48(from intermediate 10b and 1- (oxan-2-yl)pyrazole-3- carbaldehyde)

158

Preparation of Intermediate 25

A mixture of (4S)-1-Boc-4-methyl-L-proline (174 mg, 0.761 mmol), HBTU(288 mg, 0.761 mmol) and DIPEA (0.65 mL, 3.804 mmol) in DMF (7.5 ML) wasstirred for 1 h. Then, a solution of intermediate 10 (250 mg, 0.761mmol) in DMF (5 mL) was added. The reaction mixture was stirred at rtovernight. The reaction mixture was poured into iced water, basifiedwith a 10% aqueous solution of K₂CO₃ and extracted with EtOAc. Theorganic layer was washed with water, then brine, dried over MgSO₄,filtered and evaporated to dryness. The residue (490 mg) was purified bychromatography over silica gel (irregular SiOH, 40 g; mobile phase:NH₄OH/DCM/MeOH: 0.5/95/5). The pure fractions were collected andevaporated to dryness yielding 330 mg (yield 82%) of intermediate 25.

The intermediates in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 25, startingfrom the respective starting materials.

Quantity Yield Intermediate number Structure (mg) (%) Intermediate 26(from intermediate 10b and N-Boc- (2S,4S)-4-fluoropyrrolidine-2-carboxylic acid)

200 Intermediate 27 (from intermediate 11 and (R)-5- Bocazaspiro[2.4]heptane-6 carboxylic acid)

250 100

Preparation of Intermediate 104

DIPEA (0.48 mL; 2.775 mmol) was added to a solution of intermediate 10b(200 mg), 3-carboxybenzaldehyde (100 mg; 0.666 mmol) and HATU (317 mg;0.833 mmol) in DMF (10 mL) and the reaction mixture was stirred at roomtemperature for 4 hours. The reaction mixture was poured onto water andextracted with EtOAc. The organic layer was decanted, washed with water,then brine, dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (irregular SiOH,24 g; mobile phase: gradient from 0% MeOH, 100% DCM to 10% MeOH, 90%DCM). The pure fractions were collected and evaporated to drynessyielding 62 mg of intermediate 104.

Preparation of Intermediate 41

Under a N₂ flow, at rt, to a solution of intermediate 11 (250 mg, 0.76mmol) in DCM (12 mL) was added intermediate 42 (246 mg, 0.91 mmol). Themixture was stirred at room temperature for 3 h. The mixture was cooledto 5° C., NaBH(OAc)₃ (323 mg, 1.52 mmol) was added and the mixture wasstirred at rt overnight. Then, it was poured into ice water and thelayers were separated. The aqueous layer was extracted with DCM. Theorganic layers were combined, washed with brine then dried over MgSO₄,evaporated. The residue was crystallized from Et₂O and pentane. Thewhite precipitate was filtered off and dried under vacuum yielding 55 mg(yield 100%) of intermediate 41.

Preparation of Intermediate 43

Intermediate 11 (500 mg, 1.52 mmol), 2-(chloromethyl)-1,1-dimethylethylester-1H-pyrrole-1-carboxylic acid ) (493 mg, 2.28 mmol) and K₂CO₃ (1.05g, 7.61 mmol) in ACN (12 mL) were stirred at room temperature for 24 h.The reaction mixture was poured into ice water and EtOAc was added. Theorganic layer was separated, washed with brine, dried over MgSO₄,filtered and evaporated till dryness. The residue was purified bychromatography over silica gel (stationary phase: irregular SiOH 15-40μm 24 g, mobile phase: NH₄OH/DCM/MeOH: gradient from 0.1/97/3 to0.1/95/5). The pure fractions were mixed and evaporated yielding 100 mg(yield 14%) of intermediate 43.

The intermediates in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 43, startingfrom the respective starting materials.

Quantity Intermediate number Structure (mg) Yield (%) Intermediate 51(from 1H- pyrrole-4-carboxaldehydeand 3-bromopropionitrile)

 157 66 With 60° C. as reaction temperature Intermediate 42 (from 1H-pyrrole-4-carboxaldehydeand N-(2-bromoethyl) phthalimide)

1000 71 With reflux as reaction temperature Intermediate 52 (from 3-hydroxybenzaldehyde and 3- (Boc-amino)propyl bromide)

 340 99 With 75° C. as reaction temperature Intermediate 53 (from 1H-pyrazole-4-carbaldehydeand 2- bromoethoxy-t-butyl dimethylsilane)

1563 65 With reflux as reaction temperature

Preparation of Intermediate 50

Under a N₂ flow, to a solution of intermediate 11 (202 mg, 0.62 mmol) inDCM (10 mL) was added tert-butyl 4-formyl-1H-pyrazole-1-carboxylate (133mg, 0.68 mmol) and AcOH (35 μL, 0.62 mmol). The mixture was stirred atroom temperature for 2 h. NaBH(OAc)₃ (521 mg, 2.46 mmol) was added andthe mixture was stirred at rt overnight, poured into ice water and thelayers were separated. The aqueous layer was extracted with DCM. Theorganic layers were combined, washed with brine then dried over MgSO₄,evaporated. The residue was purified by chromatography over silica gel(stationary phase: irregular SiOH 15-40 μm 24 g, mobile phase: DCM/MeOH(+10% NH₄OH): 97/3). The pure fractions were mixed and evaporatedyielding 145 mg (yield 46%) of intermediate 50.

Preparation of Intermediate 55

In a sealed tube, under a N₂ flow, intermediate 53 (349 mg, 1.37 mmol)and Ti(OiPr)₄ (436 μL, 1.83 mmol) were added to a solution ofintermediate 11 (300 mg, 0.914 mmol) in THF (6 mL). The solution wasstirred at 50° C. for 5 hours then, at rt overnight. The reactionmixture was cooled to 5° C. and 2N iPrMgCl in THF (2.28 mL, 4.57 mmol)was added dropwise. The reaction mixture was allowed to rise slowly tort and stirred overnight. The reaction mixture was diluted with EtOAcand poured onto a 10% aqueous solution of K₂CO₃. The insoluble materialwas removed by filtration over Celite®. The organic layer was decanted,washed with brine, dried over MgSO₄, filtered and evaporated to dryness.The residue was purified by chromatography over silica gel (irregularSiOH, 40 g; mobile phase: MeOH/DCM: gradient from 0/100 to 10/90). Thepure fractions were collected and evaporated to dryness yielding: 0.3 g(yield 54%) of intermediate 55.

Preparation of intermediate 63

Intermediate 63a

and Intermediate 63b

A solution of tert-butyl 3-acetylazetidine-1-carboxylate (364 mg; 1.83mmol), intermediate 11 (400 mg; 1.22 mmol), titanium(IV)isopropoxide(725 μL; 2.44 mmol) in ethanol (2 mL) was stirred at 45° C. for 30 min(solution become dark yellow). Ethanol (12 mL) and NaBH₄ (138 mg; 3.66mmol) were added and the solution become yellow pale. The reactionmixture was stirred at room temperature overnight. Then, it was pouredonto a 10% aqueous solution of K₂CO₃ and DCM. The insoluble was filteredthrough a pad of Celite®. The organic layer was decanted, filteredthrough Chromabond® and the solvent was evaporated 624 mg of pale yellowoil which was purified by chromatography over silica gel (SiO₂; 25 g;mobile phase: gradient from 98% DCM, 2% MeOH to 96% DCM, 4% MeOH). Thefractions containing the product were collected and the solvent wasevaporated to give 223 mg (36%) of intermediate 63 as a white foam.Intermediate 63 was purified by chiral SFC (Stationary phase: CHIRALCELOJ-H 5 μm 250×20 mm, Mobile phase: 92% CO2, 8% MeOH (0.3% iPrNH₂)). Thepure fractions were collected and the solvent was evaporated to give 83mg (13%) of intermediate 63a as a colourless oil and 90 mg (14%) ofintermediate 63b.

The intermediate in the Table below were prepared by using an analogousmethod as described for the preparation of intermediate 63, startingfrom the respective starting materials.

Intermediate number Structure Quantity Yield Intermediate 60 (fromintermediate 11 and N-Boc-4- acetylpiperidine

600 mg 52% Intermediate 60a and intermediate 60b From chiral SFCseparation of intermediate 60: (Chiralpak AD-H 5 μm 250 * 30 mm; mobilephase: 75% CO₂, 25% iPrOH(0.3% iPrNH₂)).

221 mg 20% intermediate 60a

229 mg 20% Intermediate 60b Intermediate 109 (from intermediate 11 andintermediate 108)

324 mg 33%

Preparation of Intermediate 68

Intermediate 68a

Intermediate 68b

and Compound 61

A solution of intermediate 113 (1.67 g; 6.54 mmol) in THF (15 mL) wasadded to a solution of intermediate 11 (1.4 g; 4.36 mmol) and TFA (2 mL;26.16 mmol) in THF (30 mL). The reaction mixture was stirred at rtovernight. Then NaBH(OAc)₃ (2.77 g; 13.08 mmol) was added portionwise.The reaction mixture was stirred at rt for 10 days. The solution waspoured out into a 10% aqueous solution of K₂CO₃ and EtOAc was added. Themixture was extracted with EtOAc (3×). The organic layers were combined,washed with brine, dried over MgSO₄, filtered and the solvent wasevaporated. The residue (2.9 g; yellow oil) was purified bychromatography over silica gel (SiO₂; 40 g; eluent: from 97% DCM, 3%MeOH, 0.3% NH₄OH to 90% DCM, 10% MeOH, 1% NH₄OH). The desired fractionswere collected and the solvent was evaporated to give 266 mg ofcolourless oil intermediate 68, and 215 mg of colourless oil fraction 1.

Intermediate 68 was purified by chiral SFC (Lux-cellulose-2 5 μm 250*30mm, mobile phase: 50% CO₂, 50% MeOH (0.3% iPrNH₂)). The pure fractionswere collected and the solvent was evaporated to give 114 mg (5%) ofcolourless oil intermediate 68a and 109 mg (4%) of colourless oilintermediate 68b.

Fraction 1 was purified by reverse phase (YMC-actus Triart C18 10 μm30*150 mm, mobile phase: gradient from 65% NH₄HCO₃ 0.2%, 35% ACN to 25%NH₄HCO₃ 0.2%, 75% ACN). The fractions containing the product werecollected and the solvent was evaporated. The residue (160 mg;colourless oil) was freeze-dried with water-ACN to give 90 mg (6%) ofwhite solid compound 61.

Preparation of Intermediate 69

Intermediate 69a

and Intermediate 69b

A solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(5.07 g; 20.08 mmol), 2-BOC-2,7-diaza-spiro[4.4]nonane (5 g; 22.09 mmol)and DIPEA (6.9 mL; 40.17 mmol) in iPrOH (80 mL) was heated at 90° C.overnight. The solution was cooled to rt and the solution was pouredinto water then extracted with EtOAc (3×). The organic layer was washedwith brine, dried over MgSO₄ and the solvent was evaporated to dryness.The residue (9 g, pale brown solid) was taken up with diethylether, theprecipitate was filtered and dried to give 8.4 g of intermediate 69(95%, off-white solid). Intermediate 69 was purified by chiral SFC(Chiralpak IG 5 μm 250*20 mm, mobile phase: 65% CO₂, 35% iPrOH (0.3%iPrNH₂)). The pure fractions were collected and the solvent wasevaporated to give 4.07 g of intermediate 69a (46%, yellow foam) and4.29 g of intermediate 69b (48%, yellow foam).

Preparation of Intermediate 70a

The intermediate 70a was prepared by using an analogous method asdescribed for the alternative preparation of intermediate 11, startingfrom the respective starting material intermediate 69a.

The intermediate in the Table below were prepared by using an analogousmethod as described for the alternative preparation of intermediate 11,starting from the respective starting materials.

Intermediate number Structure Quantity Yield Intermediate 70b (fromintermediate 69b

3.4 g Quant.

Preparation of intermediate 71

Intermediate 71a

and Intermediate 71b

Method A:

A mixture of intermediate 11b (4.2 g) and intermediate 72 (1.6 g; 7mmol) in THF (50 mL) was stirred at RT overnight. Then, NaBH(OAc)₃ (3 g;14 mmol) was added portion-wise. The reaction mixture was stirred atroom temperature for 24 hours. The solution was poured onto cold water,basified with an aqueous solution of NaOH 3N and EtOAc was added. Theorganic layer was separated, dried over MgSO₄, filtered and evaporatedto dryness. The residue was purified by chromatography over silica gel(irregular SiOH 80 g, mobile phase: gradient from 98% DCM, 2% MeOH (+10%NH₄OH) to 95% DCM, 5% MeOH (+10% NH₄OH)). The pure fractions werecollected and evaporated to dryness yielding 852 mg of intermediate 71.

The enantiomers were separated by chiral SFC (CHIRALCEL OD-H 5 μm 250*30mm; mobile phase: 70% CO₂, 30% EtOH). The pure fractions were collectedand evaporated to dryness yielding 294 mg of intermediate 71a and 303 mgof intermediate 71b.

Method B:

The experiment was performed 6 times on the same quantity (640 mg; 1.95mmol) Ti(OEt)₄ (0.8 mL; 3.9 mmol) was added at room temperature to asolution of intermediate 11 (640 mg; 1.95 mmol) and intermediate 72 (665mg; 2.92 mmol) in DCE (20 mL) and MeOH (8 mL). The reaction mixture wasstirred at RT for 24 h, cooled at 10° C. then NaBH₃CN (367 mg; 5.84mmol) was added portion wise. The reaction mixture was stirred at roomtemperature for 8 days. The solutions were gathered for the work-up:poured out into cold water, basified with K₂CO₃ powder and extractedwith DCM. The suspension was filtered through a pad of Celite®. Thefiltrate was decanted, dried over MgSO₄, filtered and evaporated todryness.

The residue was purified by chromatography over silica gel (irregularSiOH, 40 g; mobile phase: gradient from 100% DCM, 0% MeOH to 97% DCM, 3%MeOH, 0.1% NH₄OH). The pure fractions were collected and evaporated todryness yielding 1.7 g (28%) of intermediate 71.

The enantiomers were separated by chiral SFC (Chiralcel OD-H 5 μm 250*30mm; mobile phase: 70% CO₂, 30% EtOH (0.3% iPrNH₂)). The pure fractionswere collected and evaporated to dryness yielding 697 mg (11%) ofintermediate 71a and 727 mg (12%) of intermediate 71b.

The intermediates in the table below were prepared following Method A asdescribed for the preparation of intermediate 71, 71a and 71b startingfrom the respective starting materials.

Intermediate number Structure Quantity Yield intermediate 88

130 mg From intermediate 11b and intermediate 89 intermediate 82

 40 mg From intermediate intermediate 11b and intermediate 83Intermediate 82a

and intermediate 82b

From chiral SFC separation of intermediate 82 (Stationary phase:CHIRALPAK AD-H 5 μm 250 * 30 mm, Mobile phase: 70% CO2, 30% iPOH(0.3%iPrNH₂))

Preparation of Intermediate 77

Intermediate 77a

and Intermediate 77b

Reaction mixture 1: In a sealed tube, a solution of intermediate 78 (2equivalents), intermediate 11 (100 mg; 0.305 mmol) and Ti(OiPr)₄ (6equivalents) in EtOH (0.2 mL) was heated at 45° C. for 1 hour. Themixture was cooled down to room temperature, diluted with EtOH (3 mL)and NaBH₄ (2 equivalents) was added. The reaction mixture was stirred atroom temperature for 4 hours indicating, according to LC/MS theformation of 60% of intermediate 77

Reaction mixture 2 and 3: The reaction was performed twice on the samequantity: In a sealed tube, a solution of intermediate 78 (2equivalents), intermediate 11 (450 mg; 1.37 mmol) and Ti(OiPr)₄ (6equivalents) in EtOH (0.9 mL) was heated at 45° C. for 1 hour. Themixture was cooled down to room temperature, diluted with EtOH (13 mL)and NaBH₄ (2 equivalents) was added. The reaction mixture was stirred atroom temperature for 18 hours.

The three reaction mixtures were diluted with EtOAc and poured onto amixture of 10% K₂CO₃ and brine. The suspension was sonicated for 30 minand filtered through a pad of Celite®. The organic layer was decanted,washed with 10% aqueous K₂CO₃, then brine, dried over MgSO₄, filteredand evaporated to dryness. The residue (2.6 g) was purified bychromatography over silica gel (irregular SiOH, 50 g; mobile phase:gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 0.7% NH₄OH, 7% MeOH, 93%DCM). The pure fractions were collected and evaporated to dryness. Theresidue (1.5 g; 89%) was purified a second time by chromatography oversilica gel (irregular SiOH, 40 g; mobile phase: 60% Heptane, 35% EtOAc,5% MeOH (+10% NH₄OH)). The fractions containing the product werecollected and evaporated to dryness yielding 980 mg (58%; 82% pure basedon LC/MS) of intermediate 77.

The impure fraction of intermediate 77 was further purified by achiralSFC (DIETHYLAMINOPROPYL 5 μm 150×30 mm; mobile phase: 90% CO₂, 10%MeOH).

The pure fractions were collected and evaporated to dryness yielding 620mg (37%) of intermediate 77.

The enantiomers were separated by chiral SFC (Lux Cellulose-2 5 μm250*30 mm; mobile phase: 50% CO₂, 50% MeOH (0.3% iPrNH₂)). The fractionscontaining the products were collected and evaporated to drynessyielding 276 mg (16%) of intermediate 77a and 269 mg (16%) ofintermediate 77b.

The intermediates in the table below were prepared using an analogousmethod as described for the preparation of intermediate 77, 77a and 77bstarting from the respective starting materials.

Intermediate number Structure Quantity Yield intermediate 80

240 mg 57% From intermediate 11 and tert-butyl3-propanoylazetidine-1-carboxylate

Preparation of Intermediate 72

Under N₂ at 5° C., iPrMgCl 2M in THF (19 mL; 38.33 mmol) was added to asolution of intermediate 73 (4.6 g; 18.83 mmol) in THF (70 mL). Thesolution was stirred at 5° C. for 30 min, allowed to slowly rise RT,stirred for 1 h then, heated at 40° C. for 5 h.

The reaction mixture was cooled to room temperature, poured out onto amixture of iced water and a saturated aqueous NH₄Cl solution, andextracted with EtOAc. The organic layer was decanted, dried over MgSO₄,filtered and evaporated to dryness yielding 4.7 g of intermediate 72(quantitative).

The intermediates in the table below were prepared using an analogousmethod as described for the preparation of intermediate 72 starting fromthe respective starting materials.

intermediate number Structure Quantity Yield intermediate 75

 25 g Quant. From intermediate 73 and cyclopropylmagnesium bromide 0.5in THF intermediate 78

 19 g 96% From intermediate 73 and Isobutylmagnesium bromide 0.4Mintermediate 83

 137 mg 49% From intermediate 84 and Isopropylmagnesium chloride 2M inTHF intermediate 105

 71 mg 16% From N-Methoxy-N- methyltetrahydrofuran-3-carboxamide andIsopropylmagnesium chloride 2M in THF Intermediate 108

 710 mg 26% From N-Methyl-N-methoxy-1-(tert-butoxycarbonyl)piperidine-4-acetamide and Isopropylmagnesium chloride 2Min THF Intermediate 111

 770 mg 40% From intermediate 73 and chloro[(4-methylphenyl)methyl]magnesium Intermediate 113

1230 mg 33% From 4-[(N-Methoxy-N- methylamino)carbonyl]-1-piperidinecarboxylic acid 1,1- dimethylethyl ester andIsopropylmagnesium chloride 2M in THF

Preparation of Intermediate 73

1-Boc-azetidine-3-carboxylic acid (5 g; 24.9 mmol) andN,O-dimethylhydroxylamiine hydrochloride (3.64 g; 37.3 mmol) were placedin a round bottom flask under N₂. DCM (75 mL) was added, followed byEDCI.HCl (7.15 g; 37.3 mmol), DMAP (155 mg; 1.27 mmol) and DIPEA (6.5mL, 37.4 mmol). The reaction mixture was stirred at RT for 16 h anddiluted with DCM (100 mL). The organic layer was washed with aqueous 1MHCl (2×50 mL), sat. NaHCO₃ solution (50 mL), and brine (50 mL). Theorganic phase was decanted, dried over MgSO₄, filtered, and evaporatedto dryness yielding 6.04 g (99%) of intermediate 73.

The intermediates in the table below were prepared using an analogousmethod as described for the preparation of intermediate 73 starting fromthe respective starting materials.

intermediate number Structure Quantity Yield intermediate 84

600 mg 97% From 1-Boc-3-fluoroazetidine-3- carboxylic acid

Preparation of Intermediate 85

Under N₂, a solution of intermediate 11 (204 mg; 0.62 mmol),intermediate 86 (217 mg; 0.81 mmol) and Ti(OEt)₄ (0.26 mL; 1.24 mmol) inDCE (7 mL) was stirred at RT overnight. NaBH₃CN (129 mg; 2 mmol) wasadded and the solution was stirred for 4 days. Water was added dropwisethen the solution was filtered through a pad of Celite®. The filtratewas separated. The organic layer was washed with water, dried overMgSO₄, filtered and evaporated till dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 97% DCM, 3% MeOH (+10% NH₄OH) to 95% DCM, 5% MeOH (+10%NH₄OH)). The pure fractions were collected and evaporated to drynessyielding 209 mg (80%) of intermediate 85.

The intermediate in the table below was prepared using an analogousmethod as described for the preparation of intermediate 85 starting fromthe respective starting materials.

intermediate number Structure Quantity Yield intermediate 87

145 mg 51% From intermediate 11 and 4-[(1-methyl-1H-pyrazol-4-yl)carbonyl]-1- piperidinecarboxylic acid 1,1-dimethylethylester

Preparation of Intermediate 86

2-bromo-1-(1-methyl-1H-pyrazol-4-yl)-ethanone (0.5 g; 2.46 mmol) in DMF(10 mL) were added to potassium phthalimide (0.46 g; 2.46 mmol). Thereaction mixture was stirred at RT for 5 h, poured into water-ice andEtOAc was added. The organic layer was separated, washed with water,brine, dried over MgSO₄, filtered and evaporated till dryness. Theresidue was purified by chromatography over silica gel (irregular SiOH15-40 μm, 24 g; mobile phase: 97% DCM, 3% MeOH (+10% NH₄OH)). The purefractions were collected and evaporated to dryness yielding 460 mg (69%)of intermediate 86.

Preparation of Intermediate 89

Under N₂, n-BuLi 1.6M in hexane (6.2 mL; 9.92 mmol) was added at −70° C.to a solution of 4-iodo-1-methyl-1H-pyrazole (1.7 g; 8.17 mmol) in THF(35 mL). The reaction mixture was stirred at −70° C. for 1 hour then, asolution of intermediate 73 (2 g; 8.19 mmol) in THF (10 mL) was addeddrop wise. The reaction mixture was stirred at −70° C. for 2 hours,allowed to warm up to room temperature and stirred overnight. Thesolution was poured out into a mixture of ice-water and a saturatedNH₄Cl solution, then EtOAc was added. The organic layer was decanted,dried over MgSO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (irregular SiOH, 50 g; mobilephase: gradient from 100% DCM, 0% MeOH to 98% DCM, 2% MeOH, 0.1% NH₄OH).The pure fractions were collected and evaporated to dryness yielding 320mg (15%) of intermediate 89

Preparation of Intermediate 92

Under N₂, HBTU (210 mg; 0.555 mmol) was added to a solution ofBOC-L-proline (; 119 mg; 0.555 mmol) and DIPEA (0.48 mL; 2.775 mmol) inDMF (10 mL). The solution was stirred for 30 min. Then, intermediate 10b(200 mg) was added and the solution was stirred at room temperature allover the weekend. Subsequently, the reaction mixture was poured intoiced water, basified with a 10% aqueous solution of K₂CO₃ and extractedwith EtOAc. The organic layer was washed by water, dried over MgSO₄,filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 0.5% NH₄OH, 5% MeOH, 95% DCM to 1% NH₄OH, 10% MeOH, 90%DCM). The pure fractions were collected and evaporated to drynessyielding 168 mg of intermediate 92.

The intermediates in the table below were prepared using an analogousmethod as described for the preparation of intermediate 92 starting fromthe respective starting materials.

intermediate number Structure Quantity Yield intermediate 93

510 mg From intermediate 10b and (2S,4R)-N-boc-4-methylpyrrolidine-2-carboxylic acid intermediate 101

150 mg From intermediate 10b and (2S,4R)-1- (tert-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid intermediate 96

283 mg 94% From intermediate 10 and cis-1-N-Boc- 4-methoxy-L-prolineintermediate 102

200 mg From intermediate 10b and (S)-5-Boc-5-Azaspiro[2.4]heptane-6-carboxylic acid intermediate 103

160 mg From intermediate 11c and (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4] heptane-6-carboxylic acid intermediate98

348 mg From intermediate 10b and (R)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4] heptane-6-carboxylic acid Intermediate99

389 mg 75% From intermediate 10 and (2S,4R)-1- (tert-butoxycarbonyl)-4-methoxypyrrolidine-2-carboxylic acid Intermediate 100

100 mg From intermediate 10b and N-BOC-4,4- difluoro-L-proline

Preparation of Intermediate 107

Under nitrogen, a solution of intermediate 11 (250 mg; 0.761 mmol),N-Boc-4-formylpiperidine (195 mg; 0.914 mmol) in THF (7 mL) was stirredat rt for 3 h. NaBH(OAc)₃ (323 mg; 1.52 mmol) was added and the mixturewas stirred at rt overnight. A 10% aqueous solution of K₂CO₃ and DCMwere added. The organic layer was separated, dried over MgSO₄, filteredand evaporated to dryness. The residue was purified by silica gelchromatography (irregular SiOH, 24 g; mobile phase: gradient from 100%DCM 0% MeOH (0% NH₄OH) to 90% DCM 10% MeOH (10% NH₄OH)). The fractionscontaining the product were mixed and evaporated to dryness yielding 383mg (96%) of intermediate 107.

Preparation of Intermediate 110

DIPEA (0.45 mL, 3.24 mmol) was added to an ice-cooled solution of2-(4-fluorophenyl) propanol (CAS[59667-20-8]) (0.25 g, 1.62 mmol) in DCM(1.4 mL) followed by methane sulfonyl chloride (0.155 mL, 1.95 mmol).The mixture was stirred overnight at rt. The mixture was diluted withDCM (20 mL) and washed with a saturated sodium bicarbonate solution (15mL). The solution was dried over MgSO₄, filtered and concentrated underreduced pressure yielding 0.377 g of intermediate 110. This product wasused without further purification in the next step.

Preparation of Intermediate 112

A solution of intermediate 111 (607 mg, 2.1 mmol) and titaniumisopropoxyde (1.25 mL, 1.37 mmol) in EtOH (4.6 mL) was added dropwise atroom temperature (over a period of 5 to 10 min) to a mixture ofintermediate 11 (459 mg, 1.4 mmol) and NaBH₃CN (264 mg, 4.2 mmol) inEtOH (9.2 mL). The mixture was stirred for 1 h at rt. The reactionmixture was diluted with DCM and poured onto a 10% aqueous solution ofK₂CO₃. The suspension was filtered over a pad of Celite®. The organiclayer was decanted, washed with brine, dried over MgSO₄, filtered andevaporated to dryness. The residue was purified by chromatography oversilica gel (irregular SiOH, 24 g; mobile phase: DCM/MeOH: gradient from100/0 to 90/10). The pure fractions were collected and evaporated todryness yielding 0.521 g (62%) of intermediate 112 (62%).

B. Preparation of the Compounds Example B1 Preparation of Compound 1

TEA (88.5 mg, 0.875 mmol) and benzaldehyde (46.4 mg, 0.44 mmol) weresuccessively added to a solution of intermediate 11 (190 mg, 0.44 mmol)in anhydrous DCM (4 mL) and the mixture was stirred at rt for 30 min.NaBH(OAc)₃ was then added (185.4 mg, 0.875 mmol) and the mixture wasstirred at rt overnight. Sat. aq. NaHCO₃ (10 mL) and DCM (10 mL) wereadded and the mixture decanted. The aqueous layer was extracted twicewith DCM (10 mL). The organic layers were combined, washed with brine(10 mL), dried over Na₂SO₄ and evaporated to give a yellow oil. Thecrude residue was purified by chromatography over silica gel (columnGemini 150*25 Sum, mobile phase: water (0.05% ammonia hydroxidev/v)/ACN: gradient from 55/45 to 25/75). The residue was thenfreeze-dried to give 65 mg of compound 1 (35% yield) as a yellow solid.

Example B2

Preparation of Compound 2

Intermediate 11 (100 mg, 0.3 mmol), 3,3,3-trifluoropropanal (51 mg, 0.46mmol) in dry DCM (3 mL) were stirred at rt for 1 h, then NaBH(OAc)₃ (129mg, 0.61 mmol) was added and the mixture was stirred at rt overnight.The mixture was poured into water then extracted with DCM, the organiclayer was dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (stationaryphase: irregular SiOH 15-40 μm 24 g, mobile phase: NH₄OH/DCM/MeOH:0.1/97/3). The product containing fractions were collected andevaporated to dryness yielding 58 mg (45%) of compound 2, which wasfreeze-dried with ACN/water 20/80 to give 45 mg of compound 2.

Preparation of Compound 13

A mixture of intermediate 11c (600 mg), isobutyraldehyde (160 mg; 2.221mmol), NaBH(OAc)₃ (1.57 g; 7.405 mmol) and Et₃N (0.64 mL; 4.443 mmol) inDCE (12 mL) was stirred at room temperature overnight. A saturatedaqueous solution of NaHCO₃ (20 mL) and DCM (20 mL) were added. Theorganic layer was decanted and the aqueous layer was extracted with DCM(20 mL*2). The combined organic layers were washed with brine (30 mL),dried over Na₂SO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (mobile phase: gradient frompetroleum ether/EtOAc from 100/0 to 0/100, then EtOAc/MeOH from 100/0 to85/15). The pure fractions were collected, evaporated to dryness andfreeze dried yielding 320 mg of compound 13.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 2, starting from therespective starting materials.

Quantity Compound number Structure (mg) Yield (%) Compound 3 (fromintermediate 7)

 80 as a hydrochloride salt Compound 4 (from intermediate 11)

126 47 Compound 5 (from intermediate 11)

 75 43 Compound 6 (from intermediate 11)

 55 26 Compound 7 (from intermediate 11)

 55 31 Compound 8 (from intermediate 11)

 75 43 Compound 9 (from intermediate 11)

 75 39 Compound 10 (from intermediate 11)

 40 29 Compound 11 (from intermediate 11)

 65 37 Compound 12 (from intermediate 11)

 80 41 Compound 13 (from intermediate 11)

 80 54 Compound 23 (from intermediate 11 and 1- methyl-1H-imidazole-5-carboxaldehyde)

 70 18 Compound 35 (from intermediate 11 and 1- methyl-1H-pyrazole-3-carbaldehyde)

155 40 Compound 69 (from intermediate 11 and phenylacetaldehyde(CAS[122-78-1]))

 60 22

Example B3

Preparation of Compound 14

Intermediate 10b (0.42 g), benzyl bromide (0.19 mL, 1.6 mmol), and K₂CO₃(0.55 g, 4.0 mmol) in ACN (20 mL) were stirred at rt overnight. Themixture was poured into water, extracted with EtOAc, the organic layerwas washed with brine, then dried over MgSO₄, filtered and evaporated todryness. The residue was purified by chromatography over silica gel(stationary phase: irregular SiOH 15-40 μm 40 g, mobile phase:NH₄OH/DCM/MeOH: 0.1/97/3). The product containing fractions werecollected and evaporated to dryness yielding 170 mg (31%) of compound14, which was crystallized from DIPE, filtered and dried to give 103 mgof compound 14.

The compounds and intermediates in the Table below were prepared byusing an analogous method as described for the preparation of compound14, starting from the respective starting materials.

Quantity Compound number Structure (mg) Yield (%) Compound 15 (fromintermediate 11)

 65 37 Compound 16 (from intermediate 8)

 57 as a hydrochloride salt (1.7HCl.1.9H₂O) Compound 17 (fromintermediate 11) (int. 11 was reacted with 1628318-10-4, followed bycleavage with TFA)

 60 32 Compound 18 (from intermediate 6)

130 49 Compound 19 (from intermediate 12)

 45 12 Compound 30 (from intermediate 11 and chloromethyltrimethylgermane)

 28 13 Compound 31 (from intermediate 11 and chloromethyltrimethylgermane)

 36 21

Example B4 Preparation of Compound 20

A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(150 mg, 0.59 mmol) prepared as described in Journal of MedicinalChemistry (2016), 59(3), 892-913, 2-benzyl 2,7-diaza-spiro-[4.4]nonane(CAS[885275-27-4]) (129 mg, 0.59 mmol) and DIPEA (0.31 mL, 1.78 mmol) inACN (15 mL) were heated at 80° C. overnight. The mixture was cooled andpoured into cooled water, the product was extracted with EtOAc, theorganic layer was dried over MgSO₄, filtered and evaporated to dryness.The residue was purified by chromatography over silica gel (stationaryphase: irregular bare silica 24 g, mobile phase: DCM/MeOH/NH₄OH:97/3/0.1). The product containing fractions were collected andevaporated to dryness yielding 200 mg (yield 78%) of compound 20(racemic mixture).

Preparation of Enantiomers 20a

and 20b

The enantiomers were separated via chiral SFC (stationary phase: LuxCellulose-4 5 μm 250*21.2 mm, mobile phase: CO₂/MeOH (0.3% iPrNH₂):70/30). The product containing fractions were collected and evaporatedto dryness yielding 80 mg (yield 31%) of a first eluted fraction F1 and81 mg (yield 31%) of a second eluted fraction F2.

F1 (80 mg; 0.185 mmol) was dissolved in acetone, at 10° C., and 4N HClin dioxane (2 eq, 0.37 mmol, 93 μL) was added followed by Et₂O. Themixture was evaporated to dryness and taken up with Et₂O, a precipitatewas filtered and dried giving 65 mg (yield 20%) of compound 20a as ahydrochloride salt (1.95HCl. 1.25H₂O. 0.19 Dioxane. 0.06 Et₂O)

F2 (81 mg, 0.187 mmol) was dissolved in acetone, at 10° C., and 4N HClin dioxane (2 eq, 0.37 mmol, 93 μL) was added followed by Et₂O. Themixture was evaporated to dryness, taken up with Et₂O, a precipitate wasfiltered and dried giving 49 mg (yield 15%) of compound 20b as ahydrochloride salt (2.0HCl. 1.8H₂O).

Example B5 Preparation of Compound 18

A mixture of intermediate 6 (222 mg, 0.63 mmol), benzyl bromide (82 μL,0.685 mmol) and K₂CO₃ (430 mg, 3.11 mmol) in ACN (20 mL) was stirred atrt overnight. The solution was poured out into cooled water, the productwas extracted with EtOAc, the organic layer was dried over MgSO₄,filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (stationary phase: irregular 15-40 μm 30g, mobile phase: DCM/MeOH/NH₄OH: gradient from 100/0/0 to 97/3/0.1). Theproduct containing fractions were collected and evaporated to drynessyielding 170 mg (yield 61%) of compound 18 (racemic mixture).

Preparation of Enantiomers Compound 21a

and Compound 21b

Compound 18 was separated into its enantiomers via chiral SFC(stationary phase: Lux Cellulose-25 μm 250*30 mm, mobile phase:CO₂/MeOH: 75/25). The product containing fractions were collected andevaporated to dryness yielding 72 mg (yield 26%) of a first elutedfraction F1 and 76 mg (yield 27%) of a second eluted fraction F2.

F1 was dissolved in acetone (3 mL), a solution of 4N HCl in dioxane (2eq, 80 μL, 0.32 mmol) was added dropwise at 10° C., Et₂O was added andafter 30 min a precipitate was filtered and dried giving 54 mg (yield16%) of compound 21a as a hydrochloride salt (1.8HCl. 1.9H₂O).

F2 was dissolved in acetone (3 mL), a solution of 4N HCl in dioxane (2eq, 85 μL, 0.34 mmol) was added dropwise at 10° C., Et₂O was added andafter 30 min a precipitate was filtered and dried giving 34 mg (yield10%) of compound 21b as a hydrochloride salt (1.8HCl.2.1H₂O).

Example B6 Preparation of Compound 22

To a solution of intermediate 11 (200 mg, 0.51 mmol) in ACN (5 mL) wasadded intermediate 16 (220 mg, 1.08 mmol) and K₂CO₃ (221.4 mg, 1.53mmol). The mixture was heated to 90° C. and stirred overnight. Water (10mL) and DCM (10 mL) were added to the reaction mixture. The organicphase was separated, the aqueous layer was extracted with DCM (10 mL).The organic layers were combined, washed with brine (10 mL), evaporatedto give a residue which was purified by chromatography over silica gel(Column: Gemini 150*25 5 u; mobile phase: water (0.05% ammonia hydroxidev/v)/CH₃CN: gradient from 42/58 to 12/88, gradient Time (min): 10; 100%B Hold Time (min): 2; flow Rate (ml/min): 25).

The desired fractions were collected and dried in vacuum to give theresidue. The residue was lyophilized to give 65 mg (yield 28%) ofcompound 22 as a light yellow solid.

Example B7 Preparation of Compound 24

Intermediate 17 (180 mg, 0.36 mmol) and 3N NaOH (0.61 mL, 1.82 mmol) inMeOH (10 mL) were stirred at rt for 1 h. The mixture was cooled to rt,poured into water and extracted with EtOAc. The organic layer was driedover MgSO₄, filtered and evaporated to dryness. The residue wasfreeze-dried with acetonitrile/water 20/80 yielding 130 mg of compound24(79% yield).

Example B8 Preparation of Compound 25

At 5° C., to a solution of intermediate 43 (100 mg, 0.2 mmol) in DCM (10mL), 4N HCl in dioxane (246 μL, 0.99 mmol) was added dropwise and themixture was stirred at rt for 15 h. The reaction was evaporated todryness. Then, the residue was taken-up with DCM, washed with NaHCO₃.The organic layer was dried over MgSO₄, filtered and evaporated todryness. The residue was purified by chromatography over silica gel(stationary phase: irregular SiOH 15-40 μm 24 g, mobile phase:NH₄OH/DCM/MeOH gradient from 0.5/95/5 to 1/90/10). The pure fractionswere collected and evaporated to dryness. The residue was freeze-driedwith acetonitrile/water 20/80 yielding 25 mg of compound 25(31% yield).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 25, starting fromthe respective starting materials.

Compound Quantity Yield number Structure (mg) (%) Compound 37 (fromintermediate 44)

250 84 Compound 112 (from intermediate 27)

 35 17

Example B9 Preparation of Compound 26

In a sealed tube,4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (0.15 g, 0.594mmol), 6-(phenylmethoxy)-2-azaspiro[3.3]heptane (0.145 g, 0.713 mmol)and DIPEA (0.205 mL, 1.19 mmol) in isopropanol (2 mL) were heated at 90°C. overnight. The solution was cooled to rt and poured into water thenextracted with EtOAc. The organic layer was washed with water, driedover MgSO₄, filtered and evaporated to dryness. The crude product wascrystallized from Et₂O and dried. The residue was purified bychromatography over silica gel (15-40 μm, 24 g, eluent: heptane/EtOAc:80/20 to 20/80). The pure fractions were mixed and the solvent wasevaporated. The residue was taken up by Et₂O, filtered and driedyielding 0.111 g of compound 26 (45% yield).

Example B10 Preparation of Compound 28

A mixture of intermediate 10b (200 mg),1-methyl-1H-pyrazole-4-carboxaldehyde (183 mg; 1.66 mmol) and AcOH (32μL; 0.555 mmol) in DCE (6 mL) was stirred at 50° C. for 2 hours. Thereaction mixture was cooled to room temperature and NaBH(OAc)₃ (353 mg;1.665 mmol) was added. The reaction mixture was stirred at roomtemperature overnight, poured onto a 10% aqueous solution of K₂CO₃ andextracted with DCM. The organic layer was decanted, dried over MgSO₄,filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 1% NH₄OH, 10% MeOH, 90%DCM). The pure fractions were collected and evaporated to drynessyielding 165 mg of compound 28 as an oil (73%). Compound 28 wasdissolved in ACN and HCl (4N in dioxane) (277 μL; 1.11 mmol) was added.The HCl salt was filtered but revealed to be too hydroscopic. Theresidue was then dissolved in DCM/MeOH and the organic layer was washedwith a 10% aqueous solution of K₂CO₃, dried over MgSO₄, filtered andevaporated to dryness. The resulting residue was dissolved in ACN andfumaric acid (47 mg; 0.404 mmol; 1 eq) was added and the solution wasallowed to stand until crystallization (overnight). The precipitate wasfiltered, washed with ACN then Et₂O and dried yielding 188 mg ofcompound 28 as the fumarate salt (1 equivalent based on 1H NMR).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 28, starting fromthe respective starting materials.

Quantity Yield Compound number Structure (mg) (%) Compound 34 (fromintermediate 10b and 1-methyl-1H-pyrazole-3- carboxaldehyde)

147 Compound 44 (from intermediate 10b and isobutyraldehyde)

 80

Example B11 Preparation of Compound 29

At 10° C., 4N HCl in dioxane (0.7 mL; 2.85 mmol) was added to a solutionof intermediate 50 (145 mg; 0.28 mmol) in ACN (7 mL). The solution wasstirred at rt overnight. The solution was evaporated to dryness. Theresidue was taken in ice water, basified with NH₄OH and DCM was added.The organic layer was separated, dried over MgSO₄, filtered andevaporated till dryness. The residue was purified by chromatography oversilica gel (stationary phase: irregular SiOH 15-40 μm 12 g, mobilephase: DCM/MeOH/NH₄OH 90/10/10). The pure fractions were collected andevaporated to dryness. The residue was freeze-dried withacetonitrile/water 20/80 yielding 0.050 g (43% yield) of compound 29.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 29, starting fromthe respective starting materials.

Compound Quantity Yield number Structure (mg) (%) Compound 40 (fromintermediate 47)

102 61 Compound 41 (from intermediate 48

72 55 Compound 88 (from intermediate 26

70 43

Example B12 Preparation of Compound 36

Under a N₂ flow, to a solution of intermediate 11 (287 mg, 0.87 mmol) inDCM (14 mL) was added 1-isopropyl-1H-pyrazole-4-carbaldehyde (133 mg,0.68 mmol) and AcOH (51 μL, 0.87 mmol). The mixture was stirred at rtfor 2 h. NaBH(OAc)₃ (742 mg, 3.5 mmol) was added and the mixture wasstirred at rt overnight. The mixture was poured into ice water and wasseparated. The aqueous layer was extracted with DCM. The organic layerwas washed with brine then, dried over MgSO₄, filtered and evaporated.The residue was purified by chromatography over silica gel (stationaryphase: irregular SiOH 15-40 μm 24 g, mobile phase: DCM/MeOH (+10%NH₄OH): gradient from 97/3 to 90/10. The pure fractions were collectedand evaporated to dryness. The residue was freeze-dried withacetonitrile/water: 20/80 yielding 0.057 g (15% yield) of compound 36.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 36, starting fromthe respective starting materials.

Compound Quantity Yield number Structure (mg) (%) Compound 76 (fromintermediate 11 and intermediate 51)

73 33

Example B13 Preparation of Compound 45

To a solution of intermediate 11c (200 mg) in dichloroethane (10 mL) wasadded 2-methylbenzaldehyde (59 mg; 0.494 mmol), NaBH(OAc)₃ (523 mg; 2.47mmol) and triethylamine (150 mg; 1.48 mmol). The mixture was stirred atroom temperature overnight and then, a saturated aqueous solution ofNaHCO₃ (10 mL) and DCM (10 mL) were added. The mixture was separated andthe aqueous layer was extracted with DCM (10 mL*2).

The organic layers were combined, washed with water (10 mL), dried overNa₂SO₄, evaporated to give 300 mg of a yellow oil which was purified bypreparative high-performance liquid chromatography (Column: Kromasil150*25 mm*10 um; Conditions: A: water (0.05% ammonia hydroxide v/v), B:MeCN, at the beginning: A (52%) and B (48%), at the end: A: (22%) and B(78%), Gradient Time (min) 8; Flow Rate(ml/min) 30.

The fractions containing the product were collected and the solvent wasevaporated under vacuum. The aqueous layer was lyophilized to dryness togive 150 mg (70%) of compound 45 as white solid.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 45, starting fromthe respective starting materials.

Quantity Yield Compound number Structure (mg) (%) Compound 46 (fromintermediate 11c and m- tolualdehyde)

190 Compound 47 (from intermediate 11c and 2- methoxy-5-methylbenzaldehyde)

120 Compound 48 (from intermediate 11c and 2,4- difluorobenzaldehyde)

150 Compound 49 (from intermediate 11c and p- tolualdehyde)

160 Compound 50 (from intermediate 11c and 2,4- dimethylbenzaldehyde)

155 Compound 54 (from intermediate 11c and 2- fluorobenzaldehyde)

166 Compound 56 (from intermediate 11c and 3- fluorobenzaldehyde)

150

Example B14 Preparation of Compound 108

A mixture of S)-5-methyl-5-azaspiro[2.4]heptane-6-carboxylic acid (94mg; 0.61 mmol), HBTU (231 mg; 0.61 mmol) and DIPEA (0.52 mL; 3.04 mmol)in DMF (5 mL) was stirred for 1 hour. Then, a solution of intermediate10b (200 mg) in DMF (5 mL) was added and the reaction mixture wasstirred at room temperature overnight. The reaction mixture was pouredinto iced water, basified with a 10% aqueous solution of K₂CO₃ andextracted with EtOAc. The organic layer was washed by H₂O, then brine,dried over MgSO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (irregular SiOH, 10 g; mobilephase: gradient from 3% MeOH, 97% DCM to 10% MeOH, 90% DCM). Thefractions containing the product were collected and evaporated todryness yielding 144 mg of an impure fraction 1. A second purificationwas performed (irregular SiOH, 40 g; mobile phase: 0.5% NH₄OH, 95% DCM,5% MeOH). The fractions containing the product were collected andevaporated to dryness yielding 43 mg of an impure fraction 2.

Fraction 2 was purified again by chromatography over silica gel(irregular SiOH, 10 g; mobile phase: gradient from 3% MeOH, 97% DCM to10% MeOH, 90% DCM). The fractions containing the product were collectedand evaporated to dryness. The resulting residue was taken up withdiisopropyl ether. The solid was filtered and dried yielding 17 mg ofcompound 108.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 108, starting fromthe respective starting materials.

Quantity Yield Compound number Structure (mg) (%) Compound 74 (fromintermediate 32 and N- isopropylethyl- enediamine)

10 7 Compound 75 (from intermediate 32 and 2- aminoethanol)

22 13

Example B18 Preparation of Compound 57

LiAlH₄ (66 mg, 1.73 mmol) was added to intermediate 54 (450 mg, 0.693mmol) in THF (12 mL). The reaction was stirred at rt for 1.5 h. Thereaction was quenched with a saturated aqueous solution of NH₄Cl,extracted with DCM and concentrated to afford a white solid. This solidwas purified by preparative high-performance liquid chromatography(column: Xtimate C18 150*25 mm*10 um, condition: water (0.05% ammoniahydroxide, v/v)/ACN: gradient from 52/48 to 42/58). To the aqueous layerwas added 0.1 mL IN HCl. The solution was freeze-dried yielding 30 mg ofcompound 57 as a yellow solid (HCl salt).

Example B19 Preparation of Compound 58

To a solution of intermediate 8 (200 mg, 0.5 mmol) in THF (10 mL) wasadded isobutyraldehyde (70 μL, 0.77 mmol) and TEA (0.37 mL, 2.63 mmol).The mixture was stirred at rt for 3 h. NaBH(OAc)₃ (317 mg, 1.5 mmol) wasadded and the solution was stirred at rt overnight. The solution waspoured out into cooled water and was basified with K₂CO₃ powder. Theproduct was extracted with EtOAc. The organic layer was dried overMgSO₄, filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (stationary phase: irregular bare silica40 g, mobile phase: NH₄OH/DCM/MeOH: 0.2/98/2). The residue was dissolvedin 5 mL of ACN, 2 eq of 4N HCl in dioxane (117 μL; 0.47 mmol) was addeddropwise at 10° C. Et₂O was added and after 30 mn, the solution wasevaporated to dryness, Et₂O was added and a precipitate was filtered anddried yielding 38 mg of compound 58 (HCl salt).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 58, starting fromthe respective starting materials.

Quantity Yield Compound number Structure (mg) (%) Compound 59 (fromintermediate 8 and 1- methyl-1H-pyrazole- 4-carbaldehyde)

73

Example B21 Preparation of Compound 62

1M TBAF in THF (0.815 mL, 0.815 mmol) was added dropwise to a solutionof intermediate 55 (0.248 g, 0.407 mmol) in Me-THF (8 mL) and thereaction mixture was stirred at rt overnight. The reaction mixture waspoured onto a 10% aqueous solution of K₂CO₃ and extracted with EtOAc.The organic layer was washed with 10% aqueous K₂CO₃ (30 mL), water (30mL) and brine (30 mL), dried over MgSO₄, filtered and evaporated todryness. The residue was purified by chromatography over silica gel (80g, 15-40 μm, eluent: DCM/MeOH: 97/3 to 88/12). The pure fractions weremixed and the solvent was evaporated yielding 0.043 g of compound 62(21% yield).

Example B22 Preparation of Compound 63

2,2-Dimethyl-tetrahydropyran-4-carbaldehyde (87 mg; 0.609 mmol) andNaBH(OAc)₃ (645 mg; 3.045 mmol) were added at rt to a solution ofintermediate 11 (200 mg; 0.609 mmol) in DCE (4 mL) and the reactionmixture was stirred overnight. The reaction mixture was diluted with DCMand poured onto a 10% aqueous solution of K₂CO₃. The organic layer wasdecanted, dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (irregular SiOH,24 g; mobile phase: NH₄OH/MeOH/DCM: gradient from 0/0/100 to 0.7/7/93).The pure fractions were collected and evaporated to dryness. The residuewas freeze-dried from water/ACN (80/20; 10 mL) yielding 155 mg ofcompound 63 (56% yield).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 63, starting fromthe respective starting materials.

Quantity Yield Compound number Structure (mg) (%) Compound 65 (fromintermediate 11 and 3-methyloxetane-3- carbaldehyde)

71 60

Example B23 Preparation of Compound 64

A mixture of intermediate 11 (150 mg, 0.457 mmol), 2,2,2-trifluoroethyltrifluoromethanesulfonate (69 μL, 0.502 mmol) and DBU (CAS[6674-22-2])(136 μL, 0.914 mmol) in DMSO (3 mL) was stirred at rt for 18 h. Thereaction mixture was poured onto water and extracted with EtOAc. Theorganic layer was washed several times with water, then brine, driedover MgSO₄, filtered and evaporated to dryness. The residue was purifiedby chromatography over silica gel (irregular SiOH, 24 g; mobile phase:NH₄OH/MeOH/DCM gradient from 0/0/100 to 0.7/7/93). The pure fractionswere collected and evaporated to dryness. The residue was purified asecond time by chromatography over silica gel (irregular SiOH, 10 g;mobile phase: EtOAc/heptane: gradient from 60/40 to 80/20). The purefractions were collected and evaporated to dryness. The residue wascrystallized from DIPE yielding, after drying under vacuum at 50° C.,100 mg of compound 64 (53% yield).

Example B24 Preparation of Compound 67

Under N₂, to a solution of intermediate 11 (100 mg; 0.31 mmol),2-(tetrahydro-2H-pyran-4-yl))acetaldehyde (48 μL; 0.37 mmol) in THF (3mL) were stirred at rt for 3 h. NaBH(OAc)₃ (129 mg; 0.61 mmol) was addedand the mixture was stirred at rt overnight. A 10% aqueous solution ofK₂CO₃ and EtOAc were added. The mixture was extracted with EtOAc(×3).The organic layers were combined, washed with brine then dried overMgSO₄, filtered and the solvent was evaporated.

The residue (136 mg) was purified by chromatography over silica gel(SiO₂, 4 g; gradient: from 95% DCM, 5% MeOH, 0.5% NH₄OH to 90% DCM, 10%MeOH, 1% NH₄OH ). The fractions containing the product were collectedand the solvent was evaporated to give 90 mg of colourless oil which wasrecrystallized with diisopropylether. The precipitate was filtered anddried to give 45 mg (34%) of compound 67 as a white solid.

Example B25 Preparation of Compound 71

In a sealed tube, under N₂, intermediate 35 (211 mg; 1.37 mmol) andTi(OiPr)₄ (436 μL; 1.83 mmol) were added to a solution of intermediate11 (300 mg; 0.914 mmol) in THF (6 mL). The solution was stirred at 50°C. for 5 hours then at rt overnight. The reaction mixture was cooled to5° C. and isopropyl magnesium chloride 2M in THF (2.28 mL; 4.57 mmol)was added dropwise. The reaction mixture was allowed to rise slowly tort and stirred overnight. The reaction mixture was diluted with EtOAcand poured onto a 10% aqueous solution of K₂CO₃. The precipitate wasremoved by filtration over Celite®. The organic layer was decanted,washed with brine, dried over MgSO₄, filtered and evaporated to dryness.The residue was purified by chromatography over silica gel (irregularSiOH, 40 g; mobile phase: gradient from 0% MeOH, 100% DCM to 10% MeOH,90% DCM). The fractions containing the product were collected andevaporated to dryness to give 0.337 g of an intermediate residue whichwas purified again by chromatography via reverse phase (stationaryphase: YMC-actus Triart C18 10 μm 30*150 mm, mobile phase: gradient from55% NH₄HCO₃ 0.2%, 45% ACN to 0% NH₄HCO₃ 0.2%, 100% ACN). The purefractions were collected and evaporated to dryness. The residue wasfreeze-dried with acetonitrile/water 20/80 to afford 120 mg (26%) ofcompound 71.

Example B26 Preparation of Compound 77

Hydrazine (36 μL, 0.92 mmol) was added to a solution of intermediate 41(110 mg, 0.18 mmol) in ethanol (5 mL). The solution was heated at 70° C.for 1 h 30. The reaction was cooled to rt, then poured into water andextracted with DCM. The organic layer was washed with brine, then driedover MgSO₄, filtered and evaporated to dryness. The residue was purifiedby chromatography over silica gel (stationary phase: irregular baresilica 40 g, mobile phase: NH₄OH/DCM/MeOH: 1/85/15). The pure fractionswere collected and evaporated to dryness. The residue was dissolved in 2mL of ACN, 3 eq of 6N HCl in iPrOH were added dropwise at 10° C. Et₂Owas added and after 30 mn, the precipitate was filtered and driedyielding 83 mg of compound 77(37% yield).

Example B28 Preparation of Compound 82

TFA (1.5 mL) was added to a solution of intermediate 25 (300 mg, 0.571mmol) in DCM (15 mL) and the reaction mixture was stirred for 18 h. Thereaction mixture was poured onto a 10% aqueous solution of K₂CO₃ andextracted with DCM. The organic layer was decanted, filtered overChromabond® and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 10 g; mobile phase:NH₄OH/MeOH/DCM: gradient from 0.3/3/97 to 1.5/15/85). The pure fractionswere collected and evaporated to dryness. The residue was crystallizedfrom DIPE and dried yielding 118 mg of compound 82 (48% yield).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 82, starting fromthe respective starting materials.

Compound Quantity Yield number Structure (mg) (%) Compound 315 (fromintermediate 109)

140 52 Compound 318 (from intermediate 112 and T = 0° C.)

200

Example B29 Preparation of Compound 84

Intermediate 11b (198 mg) and 4-fluorophenylacetone (68 μL, 0.51 mmol)in THF (5 mL) were stirred at rt overnight. Then NaBH(OAc)₃ (161 mg,0.76 mmol) was added portionwise. The mixture was stirred at rt for 24h. The solution was poured out into cooled water and basified with asolution of 3N NaOH, EtOAc was added. The organic layer was separated,dried over MgSO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (stationary phase: irregularbare silica 40 g, mobile phase: NH₄OH/DCM/MeOH: 0.1/97/3). The purefractions were collected and the solvent was evaporated under vacuum.The residue was freeze-dried with acetonitrile/water 20/80 yielding 30mg of compound 84.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 84, starting fromthe respective starting materials.

Compound Quantity Yield number Structure (mg) (%) Compound 307 (fromintermediate 11b and 1-(4- fluorophenyl)-3- methylbutan- 2-one)

46

Example B32 Preparation of Compound 133

Compound 137

and Compound 138

Ti(OEt)₄ (251 μL; 1.2 mmol) was added at room temperature to a solutionof intermediate 11 (200 mg; 0.6 mmol) and2,5,6,7-tetrahydro-2-methyl-4H-Indazol-4-one (120 mg; 0.8 mmol) indichloroethane (5 mL) and MeOH (1.5 mL). The mixture was stirred at rtfor 2 hours then NaBH₃CN (127 mg; 2 mmol) was added portionwise. Themixture was stirred at room temperature for 2 days. The solution waspoured out into cooled water and DCM was added. The mixture was basifiedwith K₂CO₃ powder, filtered through a pad of Celite®. The product wasextracted with DCM. The organic layer was dried over MgSO₄, filtered andevaporated to dryness.

The residue (361 mg) was purified by silica gel chromatography(Stationary phase: irregular bare silica 40 g, Mobile phase: 0.5% NH₄OH,95% DCM, 5% MeOH). The fraction containing the product were mixed andconcentrated to afford 120 mg (43%) of compound 133.

Chiral separation of compound 133 was performed via chiral SFC(Stationary phase: Chiralpak AD-H 5 μm 250*30 mm, Mobile phase: 70% CO₂,30% MeOH (0.3% iPrNH₂)). The fractions containing the products weremixed and concentrated to afford:

-   -   45 mg of fraction 1 which was freeze-dried with        acetonitrile/water 20/80 to give 40 mg (43%) of compound 137 as        a white powder.    -   46 mg of fraction 2 which was freeze-dried with        acetonitrile/water 20/80 to give 42 mg (46%) of compound 138 as        a white powder.

Example B33 Preparation of Compound 145

Compound 154

and Compound 155

A solution of 1-tetrahydro-2H-pyran-4-ylethanone (351 mg; 2.74 mmol),intermediate 11 (600 mg; 1.83 mmol), Ti(OiPr)₄ (870 μL; 2.92 mmol) inEtOH (3 mL) was stirred for 2 hours at 45° C. Additional EtOH (18 mL)and NaBH₄ (138 mg; 3.65 mmol) were added.

The reaction mixture was stirred at room temperature for 5 hours. Thereaction mixture was diluted with DCM and poured onto a 10% aqueoussolution of K₂CO₃. The insoluble material was removed by filtration overCelite®. The organic layer was separated, washed with water, dried overMgSO₄, filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 1% NH₄OH, 10% MeOH, 90%DCM). The fractions containing the product were collected and evaporatedto dryness yielding 487 mg (60%) of compound 145. The enantiomers ofcompound 145 were separated by chiral SFC (CHIRALPAK AD-H 5 μm 250*30mm; mobile phase: 70% CO₂, 30% mixture of EtOH/iPrOH 50/50 v/v). Thefractions containing the products were collected and evaporated todryness. The residues were freeze dried from from water/ACN (80/20; 10mL) yielding 171 mg (21%) of compound 154 and 178 mg (22%) of compound155.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 145, starting fromthe respective starting materials. The most relevant minor deviationsare indicated in the column ‘Compound number’.

Compound number Structure Quantity Yield Compound 279 (from intermediate11 and 1-(3-methyloxetan-3- yl)ethanone [1363381- 04-7] With 6 eq.Ti(iPrO)₄ and 2 eq. of NaBH₄.

243 mg 37% Compound 158 and compound 159 From SFC purification ofintermediate 158 CHIRALCEL OJ-H 5 μm 250*30 mm, mobile phase: 80% CO₂,20% EtOH (0.3% iPrNH₂))

 77 mg 12%

 80 mg 12% Compound 192 (from intermediate 11 and 1- (tetrahydro-2,6-dimethyl-2H-pyran-4- yl)-1-Propanone With 1.6 eq. of Ti(iPrO)₄ and 2 eq.of NaBH₄

 76 mg 14% Compound 280 (from intermediate 11b and 3-methyl-1-(tetrahydro- 2H-pyran-4-yl)-1- Butanone With 6 eq. of Ti(iPrO)₄and 2 eq. of NaBH₄

195 mg Compound 179 and compound 180 From SFC purification of compound280 CHIRALPAK AD- H 5 μm 250*33 mm; mobile phase: 75% CO₂, 25% MeOH(0.3% iPrNH₂))

 70 mg 10%

 69 mg 10% Compound 250 With 6 eq. of Ti(iPrO)₄ and 2 eq. of NaBH₄

369 mg 62% compound 163 and compound 164

123 mg 21%

123 mg 21% Compound 314 (From intermediate 11 and 4- Cyanophenylacetone

105 mg 15%

Preparation of Compound 312 (Diastereoisomer A (Mixture of 2 Compounds(RR and SS) or (RS and SR)) and Compound 313 (Diastereoisomer B (Mixtureof 2 Compounds (RS and SR) or (RR and SS)) Co 312: Diastereoisomer A (RRand SS) or (RS and SR) Co 313: Diastereoisomer B (RS and SR) or (RR andSS)

Reaction mixture 1: A solution of3-methyl-1-(6-oxaspiro[4.5]dec-9-yl)-1-Butanone (1.5 eq.), intermediate11 (100 mg; 0.285 mmol), Ti(OiPr)₄ (1.6 eq.) in ethanol (0.25 mL) wasstirred for 2 hours at 45° C. Ethanol (3 mL) was added and NaBH₄ (2 eq.)was added. The reaction mixture was stirred at room temperature for 18hours.

Reaction mixture 2: A solution of3-methyl-1-(6-oxaspiro[4.5]dec-9-yl)-1-Butanone (546 mg; 2.436 mmol; 2eq), intermediate 11 (400 mg; 1.22 mmol), Ti(OiPr)₄ (580 μL; 1.95 mmol)in ethanol (1 mL) was stirred for 2 hours at 45° C. Ethanol (12 mL) wasadded and NaBH₄ (92 mg; 2.436 mmol) was added. The reaction mixture wasstirred at room temperature for 18 hours.

The two reaction mixtures were gathered and diluted with EtOAc, pouredonto a 10% aqueous solution of K₂CO₃ and filtered through a pad ofCelite®. The organic layer was decanted, washed with brine, dried overMgSO₄, filtered and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 0.9% NH₄OH, 9% MeOH, 91%DCM). The desired fractions were collected and evaporated to dryness.

The residue (280 mg) was further separated by chromatography over silicagel (irregular SiOH, 24 g; mobile phase: 67% heptane, 33% EtOAc (+5%MeOH containing 10% NH₄OH)). The desired fractions were collected andevaporated to dryness yielding:

-   -   100 mg (12%) of compound 312 (eluted first; not pure enough)        which was further purified by chromatography over silica gel        (irregular SiOH, 24 g; mobile phase: gradient from 0% NH₄OH, 0%        MeOH, 100% DCM to 0.5% NH₄OH, 5% MeOH, 95% DCM).

The pure fractions were collected and evaporated to dryness. The purefractions were freeze dried from water/ACN (80/20; 10 mL) yielding 80 mg(10%) of compound 312.

-   -   110 mg (13%) of compound 313 (eluted second; not pure enough)        which was further purified by chromatography over silica gel        (irregular SiOH, 24 g; mobile phase: gradient from 0% NH₄OH, 0%        MeOH, 100% DCM to 0.5% NH₄OH, 5% MeOH, 95% DCM).

The pure fractions were collected and evaporated to dryness. The purefractions were freeze dried from water/ACN (80/20; 10 mL) yielding 70 mg(9%) of compound 313.

Example B34 Preparation of Compound 147

A mixture of intermediate 60a (216 mg; 0.4 mmol) and TFA (1 mL; 13.067mmol) in DCM (10 mL) was stirred at rt for 4 h. The reaction mixture wasdiluted with DCM and basified with a 10% aqueous solution of K₂CO₃. Theorganic layer was decanted, washed with water, filtered throughChromabond® and evaporated to dryness. The residue (200 mg) was purifiedby chromatography over silica gel (irregular SiOH, 10 g; mobile phase:85% DCM, 14% MeOH, 1% NH₄OH). The pure fractions were collected andevaporated to dryness. The residue was freeze-dried from water/ACN(80/20; 10 mL) yielding 136 mg of compound 147 (77%).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 147, starting fromthe respective starting materials.

Compound number Structure Quantity Yield Compound 146 (from intermediate60b

130 mg 71% Compound 150 (from intermediate 63a

 29 mg 45% Compound 148 (from intermediate 68

 50 mg Compound 168 (from intermediate 68a

 45 mg 50% Compound 169 (from intermediate 68b

 50 mg 58%

Example B35 Preparation of Compound 156

Compound 193

Compound 162

and Compound 157

A solution of 2-methyl-1-(tetrahydro-2H-pyran-4-yl)-1-Propanone (1.66 g;10.63 mmol) in THF (30 mL) was added to a solution of intermediate 11(2.33 g; 7.08 mmol) and TFA (3.3 mL; 42.5 mmol) in THF (45 mL). Thereaction mixture was stirred at rt overnight. Then NaBH(OAc)₃ (4.5 g;21.25 mmol) was added portionwise. The reaction mixture was stirred atrt for 7 days. The reaction mixture was stirred at rt for 3 days. Thesolution was poured out into a 10% aqueous solution of K₂CO₃, EtOAc wasadded. The mixture was extracted with EtOAc (3×). The organics layerswere combined, washed with brine, dried over MgSO₄, filtered and thesolvent was evaporated. The residue (3.9 g; yellow oil) was purified bychromatography over silica gel (SiO₂; 40 g; eluent: from 97% DCM, 3%MeOH, 0.3% NH₄OH to 90% DCM, 10% MeOH, 1% NH₄OH). The pure fractionswere collected and the solvent was evaporated to give 666 mg of palebrown solid compound 156.

Compound 156 was purified by reverse phase (YMC-actus Triart C18 10 μm30*150 mm, mobile phase: gradient from 50% NH₄HCO₃ 0.2%, 50% ACN to 0%NH₄HCO₃ 0.2%, 100% ACN). The pure fractions were collected and thesolvent was evaporated to give 66 mg of compound 157 (colourless oil)and 264 mg of compound 156 (8%; colourless oil).

50 mg of compound 156 was freeze-dried with water-ACN to give 47 mg ofcompound 156 (white solid).

Compound 157 was freeze-dried with water-ACN to give 53 mg of compound157 (2%, white solid).

Compound 156 (214 mg) was purified by chiral SFC (CHIRALPAK AD-H 5 μm250*30 mm, mobile phase: 75% CO₂, 25% EtOH (0.3% iPrNH₂)). The purefractions were collected and the solvent was evaporated to give 82 mg ofcompound 193 (colourless oil) and 82 mg of compound 162 (colourlessoil).

Compound 193 was freeze-dried with water-ACN to give 72 mg of compound193 (2%, white solid).

Compound 162 was freeze-dried with water-ACN to give 77 mg of compound162 (2%, white solid).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 193, starting fromthe respective starting materials.

Compound number Structure Quantity Yield Compound 309 as a mixture of 4diastereomers (ratio: 65/35) (from intermediate 11 and 105)

10 mg 7%

Example B36 Preparation of Compound 161

Compound 166

and Compound 167

In a sealed tube, under N₂, 1,3-dimethyl-1H-pyrazole-4-carbaldehyde (284mg; 2.28 mmol) and Ti(iPrO)₄ (727 μL; 3.05 mmol) were added to asolution of intermediate 11 (500 mg; 1.52 mmol) in THF (10 mL). Thesolution was stirred at 50° C. for 2 h. The reaction mixture was cooledto 5° C. and iPrMgCl (3.8 mL; 7.61 mmol) was added dropwise. Thereaction mixture was allowed to rise slowly to rt and stirred overnight.The reaction mixture was poured onto a 10% aqueous solution of K₂CO₃ andEtOAc. The insoluble was filtered through a pad of Celite® then, theorganic layer was decanted, dried over MgSO₄, filtered and the solventwas evaporated. The residue (866 mg, brown oil) was purified bychromatography over silica gel (SiO₂; 40 g; eluent: from 96% DCM, 4%MeOH, 0.4% NH₄OH to 93% DCM, 7% MeOH, 0.7% NH₄OH). The pure fractionswere collected and the solvent was evaporated to dryness. The residue(496 mg, yellow oil) was recrystallized with diethylether. Theprecipitate was filtered and dried to give 324 mg of compound 161 ((45%,white solid).

270 mg of compound 161 (was purified by chiral SFC (CHIRALPAK AD-H 5 μm250*30 mm, mobile phase: 70% CO₂, 30% iPOH (0.3% iPrNH₂)). The purefractions were collected and the solvent was evaporated to give 128 mgof compound 166 (18%, colourless oil) and 131 mg of compound 167 (18%,colourless oil).

Compound 166 was freeze-dried with water-ACN to give 110 mg of compound166 (15%, white solid).

Compound 167 was freeze-dried with water-ACN to give 115 mg of compound167 (16%, white solid).

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 161, starting fromthe respective starting materials. The most relevant minor deviationsare indicated in the column Yield.

Compound number Structure Quantity Yield Compound 172 (from intermediate11 and 1,5- dimetyl-1H-pyrazole-4- carbaldehyde

 26 mg  4% Compound 173 and compound 174 From SFC purification ofcompound 172 CHIRALCEL OD-H 5 μm 250*30 mm, mobile phase: 80% CO₂, 20%EtOH (0.3% iPrNH₂))

109 mg 15%

105 mg 14% Compound 186 (from intermediate 70b and 1-methyl-1H-Pyrazole-4- carboxaldehyde

 31 mg  9% Compound 188 and compound 189 From SFC purification ofcompound 186 (CHIRALPAK AD-H 5 μm 250*30 mm, mobile phase: 85% CO₂, 15%EtOH (0.3% iPrNH₂))

 53 mg 15%

 74 mg 21% Compound 281 From intermediate 70a and 1-methyl-1H-Pyrazole-4- carboxaldehyde.

250 71% Ti(OEt)₄ was used in the synthesis. Compound 190 and compound191 From SFC purification of compound 281 (CHIRALPAK AD- H 5 μm 250*33mm, mobile phase: 85% CO₂, 15% EtOH (0.3% iPrNH₂))

 77 mg 22%

 61 mg 17%

Example B38 Preparation of Compound 170

Intermediate 11b (198 mg) and 1-(3,5-difluorophenyl)propan-2-one)(86 mg;0.51 mmol) in THF (5 mL). The mixture was stirred at rt overnight. ThenNaBH(OAc)₃ (161 mg; 0.76 mmol) was added portionwise. The mixture wasstirred at rt for 24 h. The solution was poured out into cooled water,basified with a solution of NaOH 3N, EtOAc was added. The organic layerwas separated, dried over MgSO₄, filtered and evaporated to dryness. Theresidue (141 mg) was purified by chromatography over silica gel(irregular bare silica 40 g, mobile phase: 0.1% NH₄OH, 97% DCM, 3%MeOH). The pure fractions were collected and the solvent was evaporated.The residue (50 mg) was freeze-dried with ACN/water 20/80 to give 16 mgof compound 170.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 170, starting fromthe respective starting materials.

Compound number Structure Quantity Yield Compound 171 (from intermediate11b and 2- methoxyphenylacetone

 35 mg Compound 183 (from intermediate 11b (and 3- fluorophenylacetone

 38 mg Compound 182 (from intermediate 11b and 3- methyl-1-phenyl2-butanone

 55 mg Compound 185 (from intermediate 70a and Tetrahydropyran-4-carbaldehyde [50675- 18-8)

143 mg 56% Compound 187 (from intermediate 70b and Tetrahydropyran-4-carbaldehyde [50675- 18-8)

124 mg 48%

Example B39 Preparation of Compound 184

A mixture of intermediate 70a (250 mg; 0.73 mmol), isobutyraldehyde (200μL; 2.19 mmol) and AcOH (42 μL; 0.73 mmol) in DCE (8 mL) was stirred at50° C. for 3 h. The reaction mixture was cooled to rt and NaBH(OAc)₃(464 mg; 2.19 mmol) was added. The reaction mixture was stirred at rtovernight, poured onto a 10% aqueous solution of K₂CO₃ and extractedwith DCM. The organic layer was decanted, dried over MgSO₄, filtered andevaporated to dryness. The residue was purified by chromatography oversilica gel (irregular SiOH, 24 g; mobile phase: gradient from 100% DCMto 10% MeOH (+10% NH₄OH), 90% DCM). The pure fractions were collectedand evaporated to dryness. The hydrochloride salt was prepared: Theresidue (110 mg, 38%) was dissolved in ACN and HCl 4N in 1,4-dioxane (2eq.) was added. The solution was evaporated to dryness and taken upseveral times with ACN. The residue was crystallized from Et₂O yielding120 mg of compound 184 (HCl salt).

The compound in the Table below were prepared by using an analogousmethod as described for the preparation of compound 184, starting fromthe respective starting materials.

Compound number Structure Quantity Yield Compound 288 (from intermediate70b and iso- butyralde- hyde

35 mg 15%

Example B40 Preparation of Compound 289

A solution of intermediate 70a (100 mg; 0.29 mmol), p-tolualdehyde (50μL; 0.35 mmol) in dichloroethane (3 mL) was stirred at rt for 3 h.NaBH(OAc)₃ (124 mg; 0.58 mmol) was added and the mixture was stirred atrt overnight. A 10% aqueous solution of K₂CO₃ and DCM were added. Theorganic layer was decanted, filtered through Chromabond® and evaporatedto dryness. The residue was purified by chromatography over silica gel(irregular SiOH, 10 g; mobile phase: gradient from 0% NH₄OH, 0% MeOH,100% DCM to 0.5% NH₄OH, 5% MeOH, 95% DCM). The pure fractions werecollected and evaporated to dryness. The residue was freeze dried fromwater/ACN (80/20; 10 mL) yielding 73 mg (56%) of compound 289.

The compounds in the Table below were prepared by using an analogousmethod as described for the preparation of compound 289, starting fromthe respective starting materials. The most relevant minor deviationsare indicated in the column Yield.

Compound number Structure Quantity Yield Compound 290 (from intermediate70b and p- tolualdehyde)

85 mg 65 Compound 291 (from intermediate 70b and 4- fluorobenzaldehyde)

89 mg 68% THF as solvent Compound 292 (from intermediate 70a and 4-fluorobenzaldehyde)

86 mg 65% THF as solvent Compound 293 (from intermediate 70a and 3-methylbutyraldehyde)

68 mg 56% THF as solvent Compound 294 (from intermediate 70b and 3-methylbutyraldehyde)

72 mg 60% THF as solvent Compound 295 (from intermediate 70a and 1-methyl-1H-pyrazole-4- carbaldehyde)

50 mg 39% THF as solvent Compound 296 (from intermediate 70a andphenylacetaldehyde)

50 mg 38% THF as solvent Compound 297 (from intermediate 70b andphenylacetaldehyde)

18 mg 11% THF as solvent Compound 298 (from intermediate 70a and 2-(4-fluorophenyl) acetaldehyde)

20 mg 14% Compound 299 ((from intermediate 70b and 1-methyl-1H-pyrazole-4- carbaldehyde)

74 mg 58% THF as solvent

Example B41 Preparation of Compound 273

Compound 200

and Compound 222

A mixture of intermediate 71 (546 mg; 1.01 mmol) and TFA (1.5 mL; 19.6mmol) in DCM (15 mL) was stirred at RT for 4 hours. The mixture wasevaporated to dryness.

The residue was taken up with DCM and H₂O then, basified with aqueousNaOH 3N. The organic layer was decanted, dried over MgSO₄, filtered andevaporated to dryness yielding 400 mg (90%) of compound 222.

The enantiomers were separated by chiral SFC (Chiralpak AD-H 5 μm 250*30mm; mobile phase: 50% CO₂, 50% EtOH (0.3% iPrNH₂)). The pure fractionswere collected and evaporated to dryness yielding 106 mg (24%) ofcompound 273 and 130 mg (29%) of compound 200.

Compound 273 can also be prepared from intermediate 71a using the sameprocedure. Compound 200 can also be prepared from intermediate 71b usingthe same procedure. The compounds in the table below were prepared usingan analogous method as described for the preparation of compound 273starting from the respective starting materials.

compound number Structure Quantity Yield compound 211

125 mg 56% From intermediate 77a compound 213

123 mg 55% From intermediate 77b compound 259

 96 mg 49% From intermediate 80 compound 269

 18 mg 55% From intermediate 82 compound 269a

170 mg 99% From intermediate 82a compound 269b

193 mg Quant. From intermediate 82b

Example B42 Preparation of Compound 274

and Compound 203

Under N₂ at RT, 1-methyl-1H-pyrazole-4-carbaldehyde (100 mg; 0.913 mmol)and Ti(OEt)₄ (0.25 mL; 1.233 mmol) were added to a solution ofintermediate 11 (200 mg; 0.609 mmol) in THF (3 mL). The solution wasstirred at room temperature for 20 hours. The reaction mixture wascooled to 5° C. and iPrMgCl 2M in THF (1.5 mL; 3.045 mmol) was addeddropwise. The reaction mixture was stirred for 30 min at 5° C., allowedto rise slowly RT over 6 hours and poured onto a cold aqueous solutionof K₂CO₃. DCM was added and the reaction mixture was filtered through apad of Celite®. The insoluble material was washed several times withDCM. The organic layer was decanted, filtered over Chromabond® andevaporated to dryness. The residue was purified by chromatography oversilica gel (irregular SiOH, 24 g; mobile phase: gradient from 0% NH₄OH,0% MeOH, 100% DCM to 0.8% NH₄OH, 8% MeOH, 92% DCM). The pure fractionswere collected and evaporated to dryness yielding 195 mg (69%) ofracemic compound. The enantiomers were separated by chiral SFC (LuxCellulose-2 5 μm 250*30 mm; mobile phase: 55% CO₂, 45% MeOH (0.3%iPrNH₂)). The two fractions were freeze dried from water/ACN (80/20; 12mL) yielding 80 mg (28%) of compound 274 and 82 mg (29%) of compound203.

Example B43 Preparation of Compound 197

Compound 198

and Compound 199

Under N₂ at RT, 1-methyl-1H-pyrazole-4-carbaldehyde (99 mg; 0.9 mmol)and Ti(OEt)₄ (0.25 mL; 1.2 mmol) were added to a solution ofintermediate 11 (197 mg; 0.6 mmol) in THF (3 mL), the solution wasstirred at RT for 20 hours. The reaction mixture was cooled to 0° C. andCH₃MgBr (3M in Et₂; 1 mL; 3 mmol) was added dropwise. The solution wasstirred for 30 min at 0° C. and allowed to slowly rise RT for 6 hours.The solution was poured onto a mixture of cold water and aqueoussaturated NH₄Cl then EtOAc was added. The mixture was filtered through apad of Celite® and extracted with EtOAc. The organic layer was driedover MgSO₄, filtered and evaporated to dryness. The residue was purifiedby chromatography over silica gel (irregular SiOH, 24 g; mobile phase:gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 0.5% NH₄OH, 5% MeOH, 95%DCM). The pure fractions were collected and evaporated to drynessyielding 130 mg (50%) of compound 197. The enantiomers were separated bychiral SFC (Lux Cellulose-4 5 μm 250*21.2 mm; mobile phase: 60% CO₂, 40%MeOH (+0.3% iPrNH₂)). The pure fractions were collected, evaporated andfreeze dried from water/ACN (80/20) yielding 46 mg (17%) of compound 198and 45 mg (17%) of compound 199.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 274 starting from therespective starting materials.

Compound number Structure Quantity Yield compound 214

 19 mg 13% From intermediate 11, 1-Isopropyl-1H- pyrazole-4-carbaldehydeand iPrMgCl 2M in THF compound 283

143 mg 52% compound 283 From intermediate 11, 1-methyl-1H-pyrazole-4-carbaldehyde and EtMgBr 1M in THF compound 220

 58 mg 21% compound 220 and compound 221

 61 mg 22% compound 221 From purification of compound 283 by Chiral SFC:CHIRALPAK IC 5 μm 250 * 21.2 mm; mobile phase: 60% CO₂, 40% EtOH (0.3%iPrNH₂) compound 223

 28 mg 20% From intermediate 11, tert-butyl 4-formyl-1H-pyrazole-1-carboxylate and MeMgBr 3M in Et₂O compound 228

 77 mg 57% From intermediate 11, isothiazole-4- carbaldehyde and MeMgBr3M in Et₂O compound 230

343 mg 78% Compound 230 From intermediate 11, 1-methyl-1H-pyrazole-4-carbaldehyde and isobutylmagnesium bromide 2M in Et₂Ocompound 247

118 mg 27% Compound 247 and compound 248

114 mg 26% Compound 248 From purification of compound 230 by chiral SFC:Lux Cellulose-2 5 μm 250 * 30 mm; mobile phase: 55% CO₂, 45% MeOH (0.3%iPrNH₂). compound 231

 25 mg 19% From intermediate 11, isoxazole-4- carbaldehyde and MeMgBr 3Min Et₂O compound 233

 26 mg 19% From intermediate 11, pyridine-3- carboxaldehyde and MeMgBr3M in Et₂O compound 235

 23 mg  8% Compound 235 and compound 236

 17 mg  6% Compound 236 and compound 237

 27 mg  9% Compound 237 and compound 238

 29 mg 10% Compound 238 From intermediate 11, 1-methyl-1H-pyrazole-4-carbaldehyde and sec-butyl magnesium chloride 25% wt in THFchiral SFC purification: CHIRALPAK AD-H 5 μm 250 * 30 mm; mobile phase:80% CO₂, 20% iPrOH (0.6% iPrNH₂). Compounds 237 and 238 were obtainedafter an additional chiral SFC: CHIRALPAK AD-H 5 μm 250 * 30 mm; mobilephase: 80% CO₂, 20% iPrOH (0.6% iPrNH₂) then chiral SFC: Chiralcel OD-H5 μm 250 × 21.2 mm; mobile phase: 82% CO₂, 18% iPrOH (0.3% iPrNH₂).compound 284

174 mg 50% Compound 284 From intermediate 11, 1-methyl-1H-pyrazole-4-carbaldehyde and n-butyl magnesium chloride 2M in THFcompound 243

 58 mg 20% Compound 243 and compound 244

 64 mg 22% Compound 244 From chiral SFC separation of compound 384:CHIRALPAK IC 5 μm 250 * 30 mm; mobile phase: 60% CO₂, 40% EtOH (0.3%iPrNH₂). compound 285

 75 mg 22% Compound 285 From intermediate 11, tert-butyl 4-formyl-1H-pyrazole-1-carboxylate and iPrMgCl 2M in THF compound 245

 25 mg  7% compound 245 and compound 246

 24 mg  7% compound 246 From Chiral SFC separation of compound 285:CHIRALPAK AD-H 5 μm 250 * 30 mm; mobile phase: 70% CO₂, 30% EtOH (0.3%iPrNH₂) compound 249

 25 mg 19% From intermediate 11, thiazole-5- carbaldehyde and MeMgBr 3Min Et₂O compound 286

 94 mg 71% Compound 286 From intermediate 11, 1-methyl-1H-imidazole-5-carbaldehyde [CAS39021-62-0] and MeMgBr 3M in Et₂O compound253

 22 mg 16% compound 253 and compound 254

 22 mg 15% compound 254 From Chiral SFC separation of compound 286:Chiralcel OD-H 5 μm 250 × 21.2 mm; mobile phase: 70% CO₂, 30% MeOH (0.3%iPrNH₂))

Example B44 Preparation of Compound 195

Hydrazine monohydrate (34 μL; 0.86 mmol) was added to a solution ofintermediate 85 (100 mg; 0.17 mmol) in EtOH (4 mL). The solution washeated at 50° C. for 2 h30. The reaction mixture was poured into icewater and extracted with DCM. The organic layer was separated, driedover MgSO₄, filtered and evaporated till dryness. The residue waspurified by chromatography over silica gel (irregular SiOH, 12 g; mobilephase: 90% DCM, 10% MeOH (+10% NH₄OH)). The pure fractions werecollected and evaporated to dryness. The residue was taken up with Et₂Oand evaporated to dryness yielding 35 mg (45%) of compound 195.

Example B45 Preparation of Compound 287

Compound 201

and Compound 202

A mixture of intermediate 87 (145 mg; 0.24 mmol) and TFA (0.7 mL; 9.15mmol) in DCM (7 mL) was stirred at RT overnight. The reaction mixturewas evaporated to dryness. The residue was diluted with DCM and H₂Othen, basified with aqueous NaOH 3N. The organic layer was decanted,dried over MgSO₄, filtered and evaporated to dryness yielding 100 mg(83%) of compound 287.

The enantiomers were separated by chiral SFC (CHIRALPAK IC 5 μm 250×20mm; mobile phase: 50% CO₂, 50% MeOH (+2% iPrNH₂)). The fractionscontaining each enantiomer were collected, evaporated to dryness andpurified by reverse phase chromatography (YMC-actus Triart-C18 10 μm30*150 mm; mobile phase: gradient from 75% NH₄HCO₃ 0.2%, 25% ACN to 35%NH₄HCO₃ 0.2%, 65% ACN). The pure fractions were collected, evaporated todryness and freeze dried from ACN/water (20/80) yielding 21 mg (17%) ofcompound 201 and 23 mg (19%) of compound 202.

Example B46 Preparation of Compound 224

Compound 251

and Compound 252

Under N₂, a mixture of intermediate 11b (1 g) and5-acetyl-1-methylpyrazole (168 mg 1.35 mmol) in THF (15 mL) was stirredat rt overnight. Then, NaBH(OAc)₃ (718 mg; 3.4 mmol) was added portionwise. The reaction mixture was stirred at room temperature for 72 h,poured into cold water, basified with K₂CO₃ powder and extracted withEtOAc.

The organic layer was dried over MgSO₄, filtered and evaporated todryness. The residue was purified by chromatography over silica gel(irregular SiOH, 12 g; mobile phase: gradient from 100% DCM, 0% MeOH to95% DCM, 5% MeOH, 0.3% NH₄OH). The fractions were collected andevaporated to dryness. The residue (300 mg) was purified a second timeby reverse phase chromatography (YMC-actus Triart C18 10 μm 30*150 mm;mobile phase: gradient from 65% NH₄HCO₃ 0.2% aq, 35% ACN to 25% NH₄HCO₃0.2% aq, 75% ACN). The pure fractions were collected, evaporated todryness yielding 104 mg of compound 224.

Compound 224 was submitted to chiral SFC separation ((Stationary phase:Chiralcel OD-H 5 μm 250×21.2 mm, Mobile phase: 70% CO₂, 30% EtOH(0.3%iPrNH₂)).

The fractions containing the product were mixed, concentrated andfreeze-dried (ACN/water: 80/20) to afford 48 mg of compound 251 and 46mg of compound 252.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 224 starting from therespective starting materials.

compound number Structure Quantity Yield compound 232

63 mg From intermediate 11b and 4- acetylpyridine compound 258

45 mg From intermediate 11b and 5- acetylpyrimidine

Example B47 Preparation of Compound 234

A mixture of intermediate 88 (130 mg; 0.225 mmol) and HCl 4M in1,4-dioxane (0.7 mL; 2.8 mmol) in MeOH (7 mL) was stirred at rt for 24hours. The solution was cooled at 5° C. and Et₂O was added. Theprecipitate was filtered and dried yielding 111 mg of compound 234 (HClsalt).

Example B48 Preparation of Compound 270

and Compound 271

A mixture of intermediate 11 (300 mg; 0.854 mmol) and (R)-Styrene oxide(293 μL; 2.563 mmol) in EtOH (6 mL) was stirred at 60° C. for 4 hours.The reaction mixture was evaporated to dryness and the residue waspurified by chromatography over silica gel (irregular SiOH, 12 g; mobilephase: gradient from 0% NH₄OH, 0% MeOH, 100% DCM to 1% NH₄OH, 10% MeOH,90% DCM). The fractions containing the products were collected andevaporated to dryness yielding 157 mg of an intermediate residue whichwas purified by reverse phase chromatography (YMC-actus Triart C18 10 μm30*150 mm; mobile phase: gradient from 60% NH₄HCO₃ 0.2% aq, 40% ACN to40% NH₄HCO₃ 0.2% aq, 60% ACN). The fractions containing the productswere collected, evaporated to dryness and freeze dried from water/ACN(80/20; 10 mL) yielding 58 mg (15%) of compound 270 and 62 mg (16%) ofcompound 271.

Example B50 Preparation of Compound 105

TFA (2.5 mL) was added to a solution of intermediate 93 (500 mg; 0.95mmol) in DCM (25 mL) and the reaction mixture was stirred for 18 hours.The reaction mixture was poured onto a 10% aqueous solution of K₂CO₃ andextracted with DCM. The organic layer was separated, filtered overChromabond® and evaporated to dryness. The residue was purified bychromatography over silica gel (irregular SiOH, 10 g; mobile phase: 0.7%NH₄OH, 7% MeOH, 93% DCM). The pure fractions were collected andevaporated to dryness. The residue was crystallized from diisopropylethyl ether and dried yielding 120 mg (29%) of compound 105.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 105 starting from therespective starting materials.

compound or number Structure Quantity Yield compound 110

 78 mg 34% From intermediate 96 compound 109

117 mg 41% From intermediate 98 compound 111

 47 mg 15% From intermediate 99 compound 90

 58 mg 71% From intermediate 100

Example B51 Preparation of Compound 89

HCl 4N in 1,4-dioxane (0.708 mL; 2.833 mmol) was added to a solution ofintermediate 101 (150 mg; 0.283 mmol) in ACN (7.5 mL) and the reactionmixture was stirred at room temperature overnight. The reaction mixturewas poured onto a 10% aqueous solution of K₂CO₃ and extracted with DCM.The organic layer was decanted, filtered over Chromabond® and evaporatedto dryness. The residue was purified by chromatography over silica gel(irregular SiOH, 12 g; mobile phase: gradient from 0% MeOH, 100% DCM to15% MeOH, 85% DCM). The pure fractions were collected and evaporated todryness. The residue was taken up several times with Et₂O yielding 30 mg(25%) of compound 89.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 89 starting from therespective starting materials.

compound number Structure Quantity Yield compound 87

47 mg 29% From intermediate 102 compound 86

55 mg 42% From intermediate 102 Compound 311

181 mg 58% From intermediate 107

Example B56 Preparation of Compound 317

Intermediate 11 (266 mg, 0.812 mmol), intermediate 110 (377 mg, 1.62mmol) and K₂CO₃ in ACN (8 mL) were stirred overnight at 90° C. Themixture was poured into water then extracted with EtOAc. The organiclayer was dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (Stationaryphase: irregular SiOH 15-40 μm 40 g, mobile phase: DCM/MeOH: gradientfrom 100/0 to 92/8). The residue was purified by chromatography oversilica gel by reverse phase (stationary phase: YMC-actus Triart-C18 10μm 30*150 mm, mobile phase: 0.2% NH₄HCO₃/ACN: gradient from 60/40 to0/100). The fractions containing the product were collected andevaporated to dryness. The resulting residue was solubilized in ACN and2 equivalents of a 4N solution of HCl in dioxane was added. The mixturewas concentrated and, then freeze-dried with acetonitrile/water 20/80yielding 0.091 g of compound 317 (HCl salt).

Example B57 Preparation of Compound 320

To a solution of intermediate 11b (200 mg) in ethanol (5 mL) was added1-(3-methoxyphenyl)propan-2-one (200 mg, 1.218 mmol), PtO₂ (20 mg) andAcOH (2 drops). After stirring at 60° C. overnight under H2, thereaction mixture was concentrated to give a residue which was purifiedby prep-HPLC (Column: SunFire 19*250 mm 10 um, Mobile Phase A: 0.1%TFA/H₂O, B: ACN) to give 35 mg (12%) of compound 320 as yellow solid.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 320 starting from therespective starting materials.

compound number Structure Quantity Yield compound 321

44 mg From intermediate 11b and 1-(4- methoxyphenyl)propan-2-oneCompound 322

54 mg From intermediate 11b and 2- Fluorophenylacetone Compound 323

36 mg From intermediate 11b and 4- hydroxyphenylacetone Compound 324

40 mg From intermediate 11b and 1-(p- tolyl)propan-2-one

Example B57

COMPOUND 61 was synthesized together with intermediate 68, 68a and 68b.See synthesis protocol for intermediate 68, 68a and 68b.

Conversion Conversion C1 Preparation of Compound 196

A solution of compound 273 (106 mg; 0.24 mmol), aqueous formaldehyde 37%w/w (110 μL; 1.48 mmol) and MgSO₄ (580 mg; 4.83 mmol) in DCM (5 mL) wasstirred at RT for 1 hour. NaBH(OAc)₃ (614 mg; 2.9 mmol) was added andthe reaction mixture was stirred at RT for 15 hours. The solution waspoured into iced water, basified with K₂CO₃ and extracted with DCM (×2).The organic layer was washed with brine, dried over MgSO₄, filtered andevaporated to dryness. The residue was purified by chromatography oversilica gel (irregular SiOH, 10 g, mobile phase: gradient from 100% DCM,0% MeOH, 0% NH₄OH to 90% DCM, 10% MeOH, 0.5% NH₄OH). The pure fractionswere collected, evaporated to dryness and the residue was freeze driedfrom ACN/water (20/80) yielding 70 mg (64%) of compound 196.

The compounds in the table below were prepared using an analogous methodas described for the preparation of compound 196 starting from therespective starting materials.

compound number Structure Quantity Yield compound 304a

50 mg 28% From compound 269a Compound 304b

52 mg 30% From compound 269b

Conversion C2 Preparation of Compound 216

A solution of compound 200 (110 mg; 0.25 mmol) and aqueous formaldehyde37% w/w (19 μL; 0.25 mmol) in MeOH (5 mL) was stirred at RT for 3 hours.NaBH₄ (19 mg; 0.5 mmol) was added and the reaction mixture was stirredat RT for 15 hours, poured into ice water, basified with K₂CO₃ andextracted with DCM (×2). The organic layer was washed with brine thendried over MgSO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (irregular SiOH 300 g; mobilephase: gradient from 0.1% NH₄OH, 5% MeOH, 95% DCM to 1% NH₄OH, 10% MeOH,90% DCM). The pure fractions were collected, evaporated to dryness andfreeze-dried from ACN/water 20/80 yielding 15 mg (13%) of compound 216.

Conversion C3 Preparation of Compound 303

Under N₂ at 10° C., ethyl bromide (45 μL; 0.6 mmol) was added to asolution of compound 222 (150 mg; 0.34 mmol) and DIPEA (207 μL; 1.2mmol) in THF (3 mL). The solution was stirred at rt overnight, then,poured onto cooled water. The product was extracted with EtOAc. Theorganic layer was dried over MgSO₄, filtered and evaporated to dryness.The residue (164 mg) was purified by silica gel chromatography(Stationary phase: irregular bare silica 12 g, Mobile phase: Gradientfrom 100% DCM, 0% MeOH (+10% NH₄OH) to 90% DCM, 10% MeOH (+10% NH₄OH)).The fractions containing the product were mixed and concentrated toafford 77 mg of an intermediate fraction which was further purified byvia reverse phase (Stationary phase: YMC-actus Triart C18 10 μm 30*150mm, Mobile phase: Gradient from 65% NH₄HCO₃ 0.2%, 35% ACN to 25% NH₄HCO₃0.2%, 75% ACN). The fractions containing the product were mixed andconcentrated to afford 40 mg of a residue which was freeze-dried withacetonitrile/water 20/80 to give 34 mg (21%) of compound 303 as a whitepowder.

Conversion C4 Preparation of Compound 316

A solution of compound 315 (114 mg; 0.237 mmol), formaldehyde, 37% inwater (106 μL; 1.42 mmol) and MgSO₄ (568 mg) in DCM (5 mL) was stirredat rt for 1 hour. Then, NaBH(OAc)₃ (602 mg; 2.84 mmol) was added and themixture was stirred at rt overnight. The reaction mixture was dilutedwith DCM and basified with a 10% aqueous solution of K₂CO₃. The organiclayer was decanted, dried over MgSO₄, filtered and evaporated todryness. The residue was purified by chromatography over silica gel(irregular SiOH, 12 g; mobile phase: gradient from 0% NH₄OH, 0% MeOH,100% DCM to 1% NH₄OH, 10% MeOH, 90% DCM). The fractions containing theproduct were collected, evaporated to dryness and freeze dried fromwater/ACN (80/20; 10 mL) yielding 110 mg (94%) of compound 316.

Analytical Part LCMS (Liquid Chromatography/Mass Spectrometry) GeneralProcedure

The High Performance Liquid Chromatography (HPLC) measurement wasperformed using a LC pump, a diode-array (DAD) or a UV detector and acolumn as specified in the respective methods. If necessary, additionaldetectors were included (see table of methods below).

Flow from the column was brought to the Mass Spectrometer (MS) which wasconfigured with an atmospheric pressure ion source. It is within theknowledge of the skilled person to set the tune parameters (e.g.scanning range, dwell time . . . ) in order to obtain ions allowing theidentification of the compound's nominal monoisotopic molecular weight(MW). Data acquisition was performed with appropriate software.

Compounds are described by their experimental retention times (Rt) andions. If not specified differently in the table of data, the reportedmolecular ion corresponds to the [M+H]⁺ (protonated molecule) and/or[M−H]⁻ (deprotonated molecule). In case the compound was not directlyionizable the type of adduct is specified (i.e. [M+NH₄]⁺, [M+HCOO]⁻,etc. . . . ). For molecules with multiple isotopic patterns (Br, Cl),the reported value is the one obtained for the lowest isotope mass. Allresults were obtained with experimental uncertainties that are commonlyassociated with the method used.

Hereinafter, “SQD” means Single Quadrupole Detector, “RT” roomtemperature, “BEH” bridged ethylsiloxane/silica hybrid, “HSS” HighStrength Silica, “DAD” Diode Array Detector. All other abbreviationsused are as defined before.

TABLE 1a LCMS Method codes (Flow expressed in mL/min; column temperature(T) in ° C.; Run time in minutes). Method Mobile Flow- Run codeInstrument Column phase Gradient Column T time 1 Agilent: Phenomenex: A:CF₃COOH 90% A for 0.8 min, 0.8-50  10 1200- Luna-C18 0.1% in water , to20% A in DAD and (5 μm, B: CF₃COOH 3.7 min, held for MSD6110 2 × 50 mm)0.05% in 3 min, back to CH₃CN 90% A in 2 min. 2 Agilent: Phenomene x: A:CF₃COOH 100% A for 1 min, 0.8-50  10 1200- Luna-C18 0.1% in water, to40% A in 4 min, DAD and (5 μm, B: CF₃COOH to 15% A in MSD6110 2 × 50 mm)0.05% in 2.5 min, back to CH₃CN 100% A in 2 min. 3 Waters: Waters: A:95% 84.2% A for 0.343-40   6.2 Acquity BEH C18 CH₃COONH₄ 0.49 min, to10.5% UPLC ®- (1.7 μm, 7 mM/5% A in 2.18 min, held DAD and 2.1 × 100 mm)CH₃CN, for 1.94 min, back Quattro B: CH₃CN to 84.2% A in Micro ™ 0.73min, held for 0.73 min. 4 Waters: Waters: A: 95% 84.2% A to 10.5%0.343-40   6.1 Acquity ® BEH C18 CH₃COONH₄ A in 2.18 min, H-Class- (1.7μm, 7 mM/5% held for 1.96 min, DAD and 2.1 × 100 mm) CH₃CN, back to84.2% A SQD2 ™ B: CH₃CN in 0.73 min, held for 0.73 min. 5 Shimadzu:SunFire A: HCOOH 70% A for 2.0-40  2.6 LC- C18 5 μm 0.1% in water,0.4min, to 5% A MS2020- 50*4.6 mm B: HCOOH in 1.2 min, to 1 % SPD- 0.1%in A in 1.0 min. M20A and CH₃CN Alltech 3300ELSD 6 Waters ACQUITY A:HCOOH 90% A for 0.6-50  2.0 UPLC- UPLC 0.1% in water, 0.1 min, to 5% AQDa- BEH C18 B: HCOOH in 1.1 min, hold 5 PDA 1.7 μm 0.1% in % A in 0.8min. Detector 2.1*50 mm CH₃CN 7 Waters ACQUITY A: HCOOH 80% A for 1.00.4-50  9.0 UPLC- UPLC BEH 0.1% in water, min, to 5% A in QDa- C18 1.7μm B: HCOOH 7.0 min, hold 5 % PDA 2.1*100 mm 0.1% in A in 1.0 min.Detector CH₃CN

TABLE 1b LCMS data. Co. No. means compound number; R_(t) means retentiontime in min. LCMS Co. No. Rt (min) [M + H]⁺ Adduct/[M − H]⁻ Method  12.725 419.2 — 1  2 2.88 425.1 483.4 [M + CH₃COO]⁻ 3  3 2.9 447.2 — 1  42.26 421.1 479.4 [M + CH₃COO]⁻ 3  5 3.314 459 — 2  6 3.176 459 — 2  73.129 459 — 2  8 3.71 454.9 — 2  9 3.333 421 — 2  10 3.269 421 — 2  113.652 454.9 — 2  12 3.409 421 — 2  13 2.498 385 — 1  14 2.76 405 463.4[M + CH₃COO]⁻ 3  15 3.516 455 — 2  16 2.88 419.5 463.3 [M + CH₃COO]⁻ 4 17 3.074 482.9 — 1  18 2.94 447 — 1  19 3.92 461.2 519.7 [M + CH₃COO]⁻3  20a 3.15 433.1 491.4 [M + CH₃COO]⁻ 3  20b 3.15 433.1 491.4 [M +CH₃COO]⁻ 3  21a 3.66 447.4 445.4 3  21b 3.65 447.3 445.3 3  22 3.104 437— 1  23 2.11 423.5 481.3 [M + CH₃COO]⁻ 4  24 2.46 452.5 510.3 [M +CH₃COO]⁻ 4  25 2.43 — 466.3 [M + CH₃COO]⁻ 3  26 3.28 420.1 — 3  28 2.08409.1 — 3  29 2.09 409.1 — 3  30 2.17 423.2 481.4 [M + CH₃COO]⁻ 3  312.67 461 519.3 [M + CH₃COO]⁻ 3  34 2.16 409.1 467.4 [M + CH₃COO]⁻ 3  352.26 423.2 481.4 [M + CH₃COO]⁻ 3  36 2.39 451.2 509.5 [M + CH₃COO]⁻ 3 37 2.375 492 — 1  40 1.98 395.1 — 3  41 1.95 395.4 — 4  44 2.31 371.5429.2 [M + CH₃COO]⁻ 4  45 2.936 433 — 1  46 2.871 433 — 1  47 3.05 463 —1  48 2.79 545.9 — 1  49 2.901 433 — 1  50 3.019 447 — 1  54 2.66 436.9— 1  56 2.731 436.9 — 1  57 3.604 449 — 2  58 2.67 385 443.2 [M +CH₃COO]⁻ 3  59 2.2 423 481.2 [M + CH₃COO]⁻ 3  61 2.11 357 415.1 [M +CH₃COO]⁻ 3  62 2.45 495.2 — 3  63 2.55 455.2 513.4 [M + CH₃COO]⁻ 3  642.92 411 469.1 [M + CH₃COO]⁻ 3  65 2.46 413.1 471.2 [M + CH₃COO]⁻ 3  662.31 427.1 485.2 [M + CH₃COO]⁻ 3  67 2.25 441.5 499.4 [M + CH₃COO]⁻ 4 69 2.89 433.1 491.3 [M + CH₃COO]⁻ 3  71 2.75 509.2 — 3  74 2.54 547.4605.7 [M + CH₃COO]⁻ 3  75 2.56 506.3 564.5 [M + CH₃COO]⁻ 3  76 2.23462.2 520.4 [M + CH₃COO]⁻ 3  77 2.01 452.1 520.5 [M + CH₃COO]⁻ 3  821.95 426.4 484.2 [M + CH₃COO]⁻ 4  84 3.05 465.1 523.3 [M + CH₃COO]⁻ 3 88 2.13 488.4 546.3 [M + CH₃COO]⁻ 3 112 2.24 452.1 510.4 [M + CH₃COO]⁻3  86 2.20 452.2 510.5 [M + CH₃COO—] 3  87 2.13 438.2 496.6 [M +CH₃COO—] 3  89 2.14 430.1 488.3 [M + CH₃COO—] 3  90 2.32 448.1 506.4[M + CH₃COO—] 3 105 1.96 426.4 484.3 [M + CH₃COO—] 4 108 2.34 452.2510.4 [M + CH₃COO—] 3 109 2.13 438.1 496.4 [M + CH₃COO—] 3 110 1.93442.5 500.2 [M + CH₃COO—] 4 111 1.90 442.5 500.3 [M + CH₃COO—] 4 1632.42 489.1 547.4 [M + CH₃COO—] 3 164 2.43 489.2 547.4 [M + CH₃COO—] 3195 1.90 452.5 510.3 [M + CH₃COO—] 4 196 2.35 454.2 512.4 [M + CH₃COO—]3 197 2.29 437.2 495.5 [M + CH₃COO—] 3 198 2.25 437.2 495.4 [M +CH₃COO—] 3 199 2.25 437.2 495.5 [M + CH₃COO—] 3 200 2.28 440.1 498.3[M + CH₃COO—] 3 201 1.92 506.5 564.4 [M + CH₃COO—] 4 202 1.89 506.5564.5 [M + CH₃COO—] 4 203 2.70 465.2 523.4 [M + CH₃COO—] 3 211 2.40454.1 512.3 [M + CH₃COO—] 3 213 2.38 454.1 512.3 [M + CH₃COO—] 3 2142.98 493.3 551.5 [M + CH₃COO—] 3 220 2.43 451.1 509.3 [M + CH₃COO—] 3221 2.44 451.1 509.3 [M + CH₃COO—] 3 222 2.31 440.1 498.3 [M + CH₃COO—]3 223 2.20 423.0 421.1 3 224 2.51 437.1 495.3 [M + CH₃COO—] 3 228 2.68440.0 498.2 [M + CH₃COO—] 3 230 2.70 479.2 537.4 [M + CH₃COO—] 3 2312.59 424.1 482.3 [M + CH₃COO—] 3 232 2.58 434.1 492.3 [M + CH₃COO—] 3233 2.56 434.1 492.3 [M + CH₃COO—] 3 234 2.03 478.2 536.4 [M + CH₃COO—]3 235 2.91 479.2 537.4 [M + CH₃COO—] 3 236 2.91 479.2 537.4 [M +CH₃COO—] 3 237 2.89 479.3 537.5 [M + CH₃COO—] 3 238 2.89 479.2 537.5[M + CH₃COO—] 3 243 2.75 479.2 537.4 [M + CH₃COO—] 3 244 2.75 479.2537.4 [M + CH₃COO—] 3 245 2.54 451.1 449.2 3 246 2.53 451.1 449.2 3 2472.69 479.2 537.4 [M + CH₃COO—] 3 248 2.69 479.2 537.4 [M + CH₃COO—] 3249 2.59 440.0 498.2 [M + CH₃COO—] 3 250 2.42 489.1 547.3 [M + CH₃COO—]3 251 2.50 437.1 495.3 [M + CH₃COO—] 3 252 2.50 437.1 495.3 [M +CH₃COO—] 3 253 2.34 437.1 495.4 [M + CH₃COO—] 3 254 2.34 437.1 495.3[M + CH₃COO—] 3 258 2.42 435.1 493.2 [M + CH₃COO—] 3 259 2.16 426.1484.3 [M + CH₃COO—] 3 269 2.34 458.4 516.4 [M + CH₃COO—] 4 270 2.52449.4 507.4 [M + CH₃COO—] 4 271 2.48 449.4 507.4 [M + CH₃COO—] 4 2732.29 440.2 498.5 [M + CH₃COO—] 3 274 2.71 465.2 523.4 [M + CH₃COO—] 3133 2.36 463.2 521.5 [M + CH₃COO]⁻ 3 137 2.26 463.5 521.4 [M + CH₃COO]⁻4 138 2.27 463.5 521.4 [M + CH₃COO]⁻ 4 145 2.42 441.1 499.4 [M +CH₃COO]⁻ 3 147 2.11 440.1 498.2 [M + CH₃COO]⁻ 3 146 2.10 440.1 498.2[M + CH₃COO]⁻ 3 148 2.35 468.2 466.4 3 154 2.46 441.1 499.4 [M +CH₃COO]⁻ 3 155 2.44 441.1 499.3 [M + CH₃COO]⁻ 3 150 2.05 412.0 470.2[M + CH₃COO]⁻; 3 410.0 193 2.96 469.2 527.5 [M + CH₃COO]⁻; 3 467.8 1622.96 469.2 527.5 [M + CH₃COO]⁻; 3 467.8 156 2.96 469.2 527.4 [M +CH₃COO]⁻; 3 467.2 157 2.81 469.2 527.4 [M + CH₃COO]⁻; 3 467.2 158 2.60427.1 485.3 [M + CH₃COO]⁻ 3 159 2.60 427.1 485.3 [M + CH₃COO]⁻ 3 1612.76 479.2 537.4 [M + CH₃COO]⁻; 3 477.2 166 2.76 479.2 537.3 [M +CH₃COO]⁻; 3 477.2 167 2.75 479.2 537.3 [M + CH₃COO]⁻; 3 477.1 172 2.75479.2 537.4 [M + CH₃COO]⁻; 3 477.4 173 2.76 479.2 537.4 [M + CH₃COO]⁻; 3477.3 174 2.75 479.2 537.4 [M + CH₃COO]⁻; 3 477.2 168 2.40 468.2 526.4[M + CH₃COO]⁻; 3 466.3 169 2.39 468.2 526.4 [M + CH₃COO]⁻; 3 466.3 1703.20 483.1 541.3 [M + CH₃COO]⁻ 3 171 2.93 477.2 535.4 [M + CH₃COO]⁻ 3183 3.09 465.1 523.4 [M + CH₃COO]⁻ 3 182 3.48 475.2 533.4 [M + CH₃COO]⁻3 192 2.90 483.5 541.4 [M + CH₃COO]⁻ 4 179 3.03 483.2 541.5 [M +CH₃COO]⁻ 3 180 3.04 483.3 541.5 [M + CH₃COO]⁻ 3 184 2.73 399.4 457.3[M + CH₃COO]⁻ 4 288 2.74 399.1 457.2 [M + CH₃COO]⁻; 3 397.1 185 2.48441.5 499.4 [M + CH₃COO]⁻ 4 187 2.42 441.1 499.3 [M + CH₃COO]⁻; 3 439.4186 2.87 479.2 537.5 [M + CH₃COO]⁻; 3 477.2 188 2.80 479.5 537.4 [M +CH₃COO]⁻; 4 523.4 [M + HCOO]⁻ 189 2.78 479.5 537.4 [M + CH₃COO]⁻; 4523.2 [M + HCOO]⁻ 190 2.80 479.5 537.4 [M + CH₃COO]⁻ 4 191 2.79 479.5537.4 [M + CH₃COO]⁻ 4 289 3.32 447 505 [M + CH₃COO]⁻ 3 290 3.33 447 505[M + CH₃COO]⁻ 3 291 3.25 451 509 [M + CH₃COO]⁻ 3 292 3.25 451 509 [M +CH₃COO]⁻ 3 293 2.78 413 471 [M + CH₃COO]⁻ 3 294 2.77 413 471 [M +CH₃COO]⁻ 3 295 2.24 437 495 [M + CH₃COO]⁻ 3 296 3.07 447 505 [M +CH₃COO]⁻ 3 297 3.07 447 505 [M + CH₃COO]⁻ 3 298 3.11 465 523 [M +CH₃COO]⁻ 3 299 2.25 437 495 [M + CH₃COO]⁻ 3 303 2.42 468 526 [M +CH₃COO]⁻ 3 304a 2.69 472 530 [M + CH₃COO]⁻ 3 304b 2.70 472 530 [M +CH₃COO]⁻ 3 307 3.51 493 551 [M + CH₃COO]⁻ 3 309 2.92 & 455 513 [M +CH₃COO]⁻ 3 3.02 311 1.99 426 484 [M + CH₃COO]⁻ 3 312 3.76 537 595 [M +CH₃COO]⁻ 3 313 3.76 537 595 [M + CH₃COO]⁻ 3 314 2.90 472 530 [M +CH₃COO]⁻ 3 315 2.22 482 540 [M + CH₃COO]⁻ 3 316 2.30 496 554 [M +CH₃COO]⁻ 3 317 3.14 465 — 3 318 2.57 502 — 3 320 0.893 477.2 — 5 3211.300 477.4 — 6 322 6.213 465.3 — 7 323 1.203 463.3 — 6 324 0.692 461.2— 5

SFCMS-Methods General Procedure for SFC-MS Methods

The SFC measurement was performed using an Analytical Supercriticalfluid chromatography (SFC) system composed by a binary pump fordelivering carbon dioxide (CO₂) and modifier, an autosampler, a columnoven, a diode array detector equipped with a high-pressure flow cellstanding up to 400 bars. If configured with a Mass Spectrometer (MS) theflow from the column was brought to the (MS). It is within the knowledgeof the skilled person to set the tune parameters (e.g. scanning range,dwell time . . . ) in order to obtain ions allowing the identificationof the compound's nominal monoisotopic molecular weight (MW). Dataacquisition was performed with appropriate software.

TABLE 2a Analytical SFC-MS Methods (Flow expressed in mL/min; columntemperature (T) in ° C.; Run time in minutes; Backpressure (BPR) inbars; all other abbreviations used in the table below are as definedbefore) Run Method Flow time code column mobile phase gradient Col T BPR1 Phenomenex A: CO₂ 30% B 3.5 3 Luxcellulose-4 B: MeOH hold 3 35 103column (3 μm, (0.3% iPrNH₂) min, 100 × 4.6 mm) 2 Phenomenex A: CO₂ 25% B3.5 3 Luxcellulose-2 B: MeOH hold 3 35 103 column (3 μm (0.3% iPrNH₂)min, 100 × 4.6 mm) 3 Daicel A: CO₂ 30% B 3.5 3 Chiralpak ® B: hold 3 35103 AD-3 (3 μm, EtOH/iPrOH min, 100 × 4.6 mm) 50/50 (0.3% iPrNH₂) 4Daicel A: CO₂ 25% B 3.5 3 Chiralpak ® B: EtOH (0.3% hold 3 35 103 AD-3(3 μm, iPrNH₂) min, 100 × 4.6 mm) 5 Daicel A: CO₂ 30% B 3.5 3Chiralpak ® B: EtOH (0.3% hold 3 35 103 AD-3 (3 μm, iPrNH₂) min, 100 ×4.6 mm) 7 Daicel A: CO₂ 50% B 3.5 4 Chiralpak ® AD- B: EtOH hold 4 35103 3 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm) 8 Daicel A: CO₂ 20% B 3.56 Chiralpak ® AD- B: iPrOH hold 6 35 103 3 (3 μm, 100 × (0.3% iPrNH₂)min, 4.6 mm) 9 Daicel A: CO₂ 30% B 3.5 3 Chiralpak ® AD- B: iPrOH hold 335 103 3 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm) 10 Daicel A: CO₂ 40% B3.5 3 Chiralpak ® AD- B: iPrOH hold 3 35 103 3 (3 μm, 100 × (0.3%iPrNH₂) min, 4.6 mm) 11 Daicel A: CO₂ 25% B 3.5 3 Chiralpak ® B: MeOHhold 3 35 103 AD-3 (3 μm, (0.3% iPrNH₂) min, 100 × 4.6 mm) 12 Daicel A:CO₂ 30% B 3.5 3 Chiralpak ® B: MeOH hold 3 35 103 AD-3 (3 μm, (+0.3%min, 100 × 4.6 mm) iPrNH₂) 16 Phenomenex A: CO₂ 40% B 3.5 6 Luxcellulose 2 B: MeOH hold 6 35 103 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6mm) 17 Phenomenex A: CO₂ 50% B 3.5 10 Lux cellulose 2 B: MeOH hold 10 35103 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm) 18 Daicel A: CO₂ 40% B 3.5 3Chiralpak ® IC- B: EtOH (0.3% hold 3 35 103 3 (3 μm, 100 × iPrNH₂) min,4.6 mm) 20 Daicel A: CO₂ 20% B 3.5 3 Chiralcel ® OD- B: EtOH (0.3% hold3 35 103 3 (3 μm, 100 × iPrNH₂) min, 4.6 mm) 21 Daicel A: CO₂ 30% B 3.53 Chiralcel ® OD- B: EtOH (0.3% hold 3 35 103 3 (3 μm, 100 × iPrNH₂)min, 4.6 mm) 22 Daicel A: CO₂ 20% B 3.5 3 Chiralcel ® OD- B: iPrOH hold3 35 103 3 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm 23 Daicel A: CO₂ 30% B3.5 3 Chiralcel ® OD- B: MeOH hold 3 35 103 3 (3 μm, 100 × (0.3% iPrNH₂)min, 4.6 mm 24 Daicel A: CO₂ 15% B 3.5 3 Chiralcel ® OJ- B: EtOH (0.3%hold 3 35 103 3 (3 μm, 100 × iPrNH₂) min, 4.6 mm 25 Daicel A: CO₂ 10% B3.5 3 Chiralcel ® OJ- B: MeOH hold 3 35 103 3 (3 μm, 100 × (0.3% iPrNH₂)min, 4.6 mm 26 Daicel A: CO₂ 30% B 3.5 3 Chiralcel ® OJ- B: MeOH hold 335 103 3 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm 37 Daicel A: CO₂ 40% B3.5 3 Chiralpak ® IC- B: iPrOH hold 3 35 103 3 (3 μm, 100 × (0.3%iPrNH₂) min, 4.6 mm) 38 Daicel A: CO₂ 20% B 3.5 6 Chiralpak ® IC- B:MeOH hold 6 35 103 3 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm) 39 DaicelA: CO₂ 15% B 3.5 3 Chiralcel ® OD- B: MeOH hold 3 35 103 3 (3 μm, 100 ×(0.3% iPrNH2) min, 4.6 mm 40 Phenomenex A: CO₂ 40% B 3.5 3 Lux cellulose2 B: EtOH hold 3 35 103 (3 μm, 100 × (0.3% iPrNH₂) min, 4.6 mm)

TABLE 2b SFC-MS data. (Isomer elution order ‘A’ elutes before ‘B’ underthe described conditions) Isomer Co. R_(t) UV % elution SFCMS No. (min)Area order Method  20a 2.62 100 A 1  20b 3.00 99.71 B 1  21a 2.66 98.09A 2  21b 3.01 96.93 B 2  82 1.49 99.72 A 5 105 1.65 97.6 B 5 110 1.3297.6 A 12 111 2.27 100 B 12 163 0.94 100 A 26 164 1.44 100 B 26 198 1.54100 A 1 199 2.29 100 B 1 273 0.75 99.6 A 7 200 1.19 99.6 B 7 274 1.29100 A 2 203 2.04 100 B 2 211 1.58 100 A 12 213 2.22 99.05 B 12 220 1.29100 A 18 221 1.90 100 B 18 235* 2.81 100 A 8 236* 3.46 100 B 8 237* 1.97100 A 22 238* 2.43 100 B 22 243 1.26 100 A 18 244 1.80 99.6 B 18 2452.02 100 A 4 246 2.54 99.5 B 4 247 1.11 99.95 A 16 248 3.20 100 B 16 2510.78 100 A 21 252 1.23 99.95 B 21 253 0.99 100 A 23 254 1.82 99.2 B 23137 0.98 100 A 12 138 1.33 99.81 B 12 147 1.99 100 B 5 146 1.46 99.18 A5 154 1.15 100 A 3 155 1.35 99.14 B 3 193 1.16 100 A 4 162 1.44 99.04 B4 158 1.16 100 A 24 159 1.66 100 B 24 166 1.07 100 A 9 167 1.31 99.03 B9 173 1.22 100 A 20 174 1.81 100 B 20 168 2.15 99.17 B 10 169 1.21 100 A10 179 0.90 100 A 11 180 1.29 99.58 B 11 184 1.20 100 A 37 288 1.4098.50 B 37 185 1.34 100 A 25 187 1.58 100 B 25 295 1.04 100 A 5 299 1.2998.11 B 5 296 1.40 100 A 9 297 1.88 100 B 9 290 1.42 100 A 37 289 1.57100 B 37 291 3.50 99.92 A 38 292 3.79 100 B 38 293 1.24 100 A 39 2941.58 100 B 39 304a 1.40 100 A 40 304b 1.92 99.02 B 40 *Isomer elutionorder Co. No. 235 vs Co. No. 236; Co. No. 237 vs Co. No. 238. SFC-MS wasalso measured for compounds 188, 189, 190 and 191 under the same SFCMSconditions. The results are shown in Table 2c. (Isomer elution order ‘A’before ‘B’, ‘B’ before ‘C’, ‘C’ before ‘D’)

Isomer Co. R_(t) UV % elution SFCMS No. (min) Area order Method 188 2.78100 B 8 189 3.02 96.66 C 8 190 2.40 100 A 8 191 3.21 99.15 D 8

Optical Rotation (OR)

Optical Rotation is measured with a polarimeter 341 Perkin Elmer. Thepolarized light is passed through a sample with a path length of 1decimeter and a sample concentration of 0.2 to 0.4 gram per 100milliliters. 2 to 4 mg of the product in vial are weight, then dissolvedwith 1 to 1.2 ml of spectroscopy solvent (DMF for example). The cell isfilled with the solution and put into the polarimeter at a temperatureof 20° C. The OR is read with 0.004° of precision.

weight in gram×100/volume in ml  Calculation of the concentration:

[α]d ²⁰: (read rotation×100)/(1.000 dm×concentration).  Specificrotation (OR):

^(d) is sodium D line (589 nanometer).

TABLE 3 OR data: wavelength: 589 nm (specicied if different); solvent:DMF (specicied if different); temperature: 20° C.; ‘conc’ meansconcentration (g/100 mL); ‘OR’ means optical rotation. Co. WavelengthNo. OR (°) Conc. (nm) Solvent  20a −31.43 0.28 589 DMF  20b +26.25 0.32589 DMF  21a −38.48 0.33 589 DMF  21b +42.02 0.326 589 DMF 112 +23.110.251 589 DMF  86 −29.55 0.264 589 DMF  87 −26.22 0.267 589 DMF 108−29.61 0.206 589 DMF 109 +25.37 0.272 589 DMF 198 −28.25 0.308 589 DMF199 +20.91 0.263 589 DMF 196 +8.92 0.269 589 DMF 200 −11.54 0.26 589 DMF201 +7.81 0.269 589 DMF 202 −5.38 0.26 589 DMF 203 −6.22 0.225 365 DMF211 +5.65 0.248 589 DMF 213 −9.89 0.263 589 DMF 216 −19.57 0.281 589 DMF220 −22.63 0.243 589 DMF 221 +11.58 0.259 589 DMF 235 −12.02 0.258 589DMF 236 +8.58 0.233 589 DMF 237 +31.58 0.228 365 DMF 238 −32.09 0.215365 DMF 243 −30.12 0.332 365 DMF 244 +20.94 0.277 365 DMF 245 −17.120.292 589 DMF 247 −17.25 0.255 365 DMF 248 +5.86 0.239 365 DMF 251 −20.60.267 589 DMF 252 +12 0.275 589 DMF 253 −25.4 0.252 589 DMF 254 +17.920.318 589 DMF 273 +10.38 0.26 589 DMF 274 −5.41 0.222 589 DMF 137 +29.320.249 589 DMF 138 −24.9 0.261 589 DMF 147 +8.06 0.273 589 DMF 146 −12.650.245 589 DMF 154 −8.59 0.256 589 DMF 155 +6.64 0.241 589 DMF 150 +14.10.234 365 DMF 193 −8.62 0.232 365 DMF 162 −5.45 0.275 589 DMF 158 +16.540.26 365 DMF 159 −22.31 0.251 365 DMF 166 −12.6 0.254 589 DMF 167 +7.970.276 589 DMF 173 +17.92 0.279 365 DMF 174 −21.99 0.282 365 DMF 168−4.59 0.283 365 DMF 179 +20.19 0.208 365 DMF 180 −93.02 0.215 365 DMF184 +17.31 0.219 589 DMF 288 −26.63 0.338 589 MeOH 185 +32.45 0.256 589DMF 187 −32.45 0.228 589 DMF 289 +36.73 0.245 589 DMF 290 −41.87 0.246589 DMF 291 −37.05 0.278 589 DMF 292 +32.39 0.247 589 DMF 293 +29.150.295 589 DMF 294 −30.83 0.253 589 DMF 295 +27.38 0.263 589 DMF 296+50.44 0.113 589 DMF 297 −32.45 0.265 589 DMF 299 −33.79 0.29 589 DMF304a +8.42 0.285 589 DMF 304b −6.49 0.385 589 DMF

NMR

NMR experiments were carried out using a Bruker Avance 500 spectrometerequipped with a Bruker 5 mm BBFO probe head with z gradients andoperating at 500 MHz for the proton and 125 MHz for carbon, or using aBruker Avance DRX 400 spectrometer using internal deuterium lock andequipped with reverse double-resonance (¹H, ¹¹C, SEI) probe head with zgradients and operating at 400 MHz for the proton and 100 MHz forcarbon. Chemical shifts (δ) are reported in parts per million (ppm). Jvalues are expressed in Hz. Alternatively, some NMR experiments werecarried out using a Bruker Avance III 400 spectrometer at ambienttemperature (298.6 K), using internal deuterium lock and equipped with 5mm PABBO BB-probe head with z gradients and operating at 400 MHz for theproton and 100 MHz for carbon. Chemical shifts (S) are reported in partsper million (ppm). J values are expressed in Hz.

Compound 13:

1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.42 (s, 1H), 7.38 (s, 1H), 3.95(br s, 2H), 3.83 (br t, J=6.6 Hz, 2H), 3.63 (q, J=10.1 Hz, 2H), 3.27 (brs, 4H), 2.32 (d, J=6.8 Hz, 2H), 2.28-2.18 (m, 2H), 1.69-1.57 (m, 1H),0.90 (d, J=6.6 Hz, 6H)

Compound 20b:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.64 (br s, 1H) 8.49 (s, 1H) 7.77 (s,1H) 7.67 (br d, J=3.8 Hz, 2H) 7.35-7.51 (m, 3H) 4.32-4.48 (m, 2H)4.03-4.18 (m, 2H) 3.91 (br s, 4H) 3.17-3.61 (m, 4H) 2.00-2.32 (m, 4H)

Compound 82:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.35 (s, 1H) 7.39 (s, 1H) 4.28-4.77 (m,6H) 4.01-4.19 (m, 4H) 3.62 (br t, J=7.9 Hz, 1H) 2.88 (dd, J=9.9, 6.8 Hz,1H) 2.45 (br dd, J=9.9, 7.7 Hz, 1H) 2.00-2.21 (m, 2H) 1.17-1.28 (m, 1H)0.95 (d, J=6.3 Hz, 3H)

Compound 84:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.33 (s, 1H) 7.72 (s, 1H) 7.22 (dd,J=8.4, 5.8 Hz, 2H) 7.09 (t, J=9.0 Hz, 2H) 4.08 (q, J=11.0 Hz, 2H)3.66-3.96 (m, 4H) 3.17 (s, 4H) 2.65-2.73 (m, 1H) 2.38-2.45 (m, 1H) 2.27(dd, J=12.9, 8.5 Hz, 1H) 2.14 (br s, 2H) 0.72 (d, J=6.3 Hz, 3H)

Compound 193:

¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.32 (s, 1H) 7.70 (s, 1H) 4.07 (q,J=11.1 Hz, 2H) 3.65-3.91 (m, 6H) 3.07-3.30 (m, 6H) 2.14 (br s, 2H) 1.85(br s, 1H) 1.70-1.82 (m, 1H) 1.63 (br s, 1H) 1.28-1.52 (m, 4H) 0.88 (dd,J=9.6, 7.1 Hz, 6H)

Compound 196:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.32 (s, 1H) 7.70 (s, 1H) 4.07 (q,J=10.9 Hz, 2H) 3.52-3.97 (m, 4H) 3.34-3.43 (m, 2H) 3.02-3.22 (m, 4H)2.72 (dd, J=8.5, 6.3 Hz, 1H) 2.58 (dd, J=8.8, 6.6 Hz, 1H) 2.51-2.53 (m,1H) 2.28-2.35 (m, 1H) 2.19 (dd, J=9.5, 2.2 Hz, 1H) 2.12 (s, 4H) 1.58(td, J=7.1, 2.2 Hz, 1H) 0.79 (dd, J=14.8, 6.9 Hz, 6H)

Compound 273:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.32 (s, 1H) 7.67 (s, 1H) 4.03 (q,J=11.0 Hz, 2H) 3.87 (s, 2H) 3.79 (t, J=6.9 Hz, 2H) 3.61 (br t, J=7.6 Hz,1H) 3.46 (br t, J=7.9 Hz, 1H) 3.12-3.34 (m, 7H) 2.66-2.77 (m, 1H) 2.28(dd, J=9.1, 2.2 Hz, 1H) 2.15 (t, J=6.9 Hz, 2H) 1.62 (td, J=6.9, 2.2 Hz,1H) 0.82 (dd, J=14.2, 6.9 Hz, 6H)

Compound 274:

¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.31 (s, 1H) 7.69 (s, 1H) 7.45 (s, 1H)7.20 (s, 1H) 4.07 (q, J=10.9 Hz, 2H) 3.58-3.93 (m, 7H) 2.97-3.08 (m, 4H)2.52-2.55 (m, 1H) 2.12 (br s, 2H) 1.7 (td, J=6.6, 4.4 Hz, 1H) 0.70 (dd,J=12.6, 6.6 Hz, 6H)

Compound 320:

1HNMR (400 MHz, CD₃OD) δ 8.37 (s, 1H), 7.63 (s, 1H), 7.28 (t, J=8.8 Hz,1H), 6.87-6.86 (m, 3H), 4.30-4.18 (m, 5H), 4.08-3.93 (m, 3H), 3.91-3.88(m, 2H), 3.88-3.85 (s, 3H), 3.72-3.70 (m, 1H), 3.06 (dd, J=5.2 Hz,J=13.6 Hz, 1H), 2.64 (dd, J=9.2 Hz, J=13.6 Hz, 1H), 2.47-2.385 (m, 2H),1.19-1.18 (d, J=6.8 Hz 3H)

Compound 321:

¹H NMR (400 MHz, DMSO) δ 8.38 (d, J=5.2 Hz 1H), 7.65 (s, 1H), 7.21 (d,J=8.4 Hz, 2H), 6.92 (d, J=8.4 Hz, 2H), 3.75-4.33 (m, 11H), 3.74 (s, 3H),3.61 (s, 1H), 2.93 (dd, J=3.6 Hz, 12.8 Hz, 1H), 2.27-2.36 (m, 2H, ),1.03 (d, J=5.6 Hz, 3H).

Compound 322:

¹H NMR (400 MHz, CD₃OD) δ 8.40 (s, 1H), 7.66 (s, 1H), 7.38-7.34 (m, 2H),7.22-1.13 (m, 2H), 4.37-4.26 (m, 5H), 4.13 (s, 1H), 4.01 (s, 2H), 3.91(q, J=10.4 Hz, 2H), 3.74-3.69 (m, 1H), 2.98 (dd, J=3.6 Hz, 13.2 Hz 1H),2.68 (dd, J=9.6 Hz, 13.2 Hz 1H), 2.47 (s, 2H), 1.20 (d, J=6.4 Hz 3H).

Compound 323:

¹HNMR (400 MHz, CD₃OD) δ 8.38 (s, 1H), 7.64 (s, 1H), 7.10 (d, J=8.4 Hz,2H), 6.78 (d, J=8.4 Hz, 2H), 3.86-7.27 (m, 10H), 3.60-6.62 (m, 1H), 2.96(dd, J=4.8 Hz, 14.0 Hz, 1H), 2.60 (dd, J=4.8 Hz, 14.0 Hz 1H), 2.37-2.46(m, 2H), 1.18 (d, J=6.8 Hz, 3H).

Compound 324:

1H NMR (400 MHz, CD₃OD) δ 8.36 (s, 1H), 7.62 (s, 1H), 7.19-7.15 (m, 4H),4.28-4.16 (m, 5H), 4.14-3.98 (m, 4H), 3.93-3.87 (m, 2H), 3.66-3.64 (m,1H), 3.04 (dd, J=4.8 Hz, J=13.6 Hz, 1H), 2.63 (dd, J=9.2 Hz, J=13.6 Hz1H), 2.46-2.32 (m, 5H), 1.18 (d, J=6.4 Hz, 3H).

Pharmacological Part 1) Menin/MLL Fluorescence Polarization Assay

To a non-surface binding, black 384-well microtiter plate was added 50nL 160× test compound in DMSO and 4 μL 2× menin in assay buffer (40 mMTris HCl, pH 7.5, 50 mM NaCl, 1 mM DTT and 0.001% Tween 20). Afterincubation of test compound and menin for 10 min at ambient temperature,4 μL 2× FITC-MBM1 peptide (FITC-β-alanine-SARWRFPARPGT-NH₂) in assaybuffer was added, the microtiter plate centrifuged at 1000 rpm for 1 minand the assay mixtures incubated for 15 min at ambient temperature. Therelative amount of menin FITC-MBM1 complex present in an assay mixtureis determined by measuring the fluorescence polarization (FP) of theFITC label with a BMG Pherastar plate reader (ex. 485 nm/em. 520 nm) atambient temperature. The final concentrations of reagents in the bindingassay are 100 nM menin, 5 nM FITC-MBM1 peptide and 0.625% DMSO in assaybuffer. Dose-response titrations of test compounds are conducted usingan 11 point, three-fold serial dilution scheme, starting at 31 μM.

Compound potencies were determined by first calculating % inhibition ateach compound concentration according to equation 1:

% inhibition=((HC−LC)−(FP ^(compound) −LC))/(HC−LC))*100  (Eqn 1)

Where LC and HC are the FP values of the assay in the presence orabsence of a saturating concentration of a compound that competes withFITC-MBM1 for binding to menin, and FP^(compound) is the measured FPvalue in the presence of the test compound. HC and LC FP valuesrepresent an average of at least 16 replicates per plate. For each testcompound, % inhibition values were plotted vs. the logarithm of the testcompound concentration, and the IC₅₀ value derived from fitting thesedata to equation 2:

% inhibition=Bottom+(Top−Bottom)/(1+10{circumflex over ( )}((logIC₅₀−log[cmpd])*h))  (Eqn 2)

Where Bottom and Top are the lower and upper asymptotes of thedose-response curve, respectively, IC₅₀ is the concentration of compoundthat yields 50% inhibition of signal and h is the Hill coefficient.

2) Proliferation Assay

The anti-proliferative effect of menin/MLL protein/protein interactioninhibitor test compounds was assessed in human leukemia cell lines. Thecell lines MV-4-11 and MOLM14 harbor MLL translocations and express theMLL fusion proteins MLL-AF4 and MLL-AF9, respectively, as well as thewildtype protein from the second allele. Therefore, the MLL rearrangedcell lines MV-4-11 and MOLM14 exhibit stem cell-like HOXA/MEIS1 geneexpression signatures. K562 and KG1 were used as a control cell linescontaining two MLL wildtype alleles in order to exclude compounds thatdisplay general cytotoxic effects.

MV-4-11 and MOLM14 were cultured in RPMI-1640 (Sigma Aldrich)supplemented with 10% fetal bovine serum (HyClone), 2 mM L-glutamine(Sigma Aldrich) and 50 μg/ml gentamycin (Gibco). K562 were propagated inRPMI-1640 (Sigma Aldrich) supplemented with 20% fetal bovine serum(HyClone), 2 mM L-glutamine (Sigma Aldrich) and 50 μg/ml gentamycin(Gibco). KG1 were cultured in Iscove's MDM (Gibco) supplemented with 20%fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich) and 50μg/ml gentamycin (Gibco). Cells were kept at 0.3-2.5 million cells perml during culturing and passage numbers did not exceed 25.

In order to assess the anti-proliferative effects, 1,500 MV-4-11, 300MOLM14, 750 K562 or 1,300 KG1 cells were seeded in 200 μl media per wellin 96-well round bottom, ultra-low attachment plates (Costar, cataloguenumber 7007). Cell seeding numbers were chosen based on growth curves toensure linear growth throughout the experiment. Test compounds wereadded at different concentrations and the DMSO content was normalized to0.3%. Cells were incubated for 8 d at 37° C. and 5% CO₂. Spheroid likegrowth was monitored in real-time by live-cell imaging (IncuCyteZOOM,Essenbio, 4× objective) acquiring one image every four hours for 8 d.Confluence (%) as a measure of spheroid size was determined using anintegrated analysis tool.

In order to determine the cumulative effect of the test compounds overtime, the area under the curve (AUC) in a plot of confluence againsttime was calculated. Confluence at the beginning of the experiment (t=0)was used as baseline for the AUC calculation.

Absolute IC₅₀ values were calculated according to the followingprocedure:

% Control=(AUC sample/AUC control)*100

AUC control=mean AUC of control values (cells without compound/DMSO asvehicle control)

A non-linear curve fit was applied using the least squares (ordinary)fit method to the plot of % control versus compound concentration. Basedon this, the absolute IC₅₀ value (half maximal inhibitory concentrationof the test compound causing an anti-proliferative effect of 50%relative to the vehicle control) was calculated.

3) Menin/MLL Homogenous Time-Resolved Fluorescence (HTRF) Assay

To an untreated, white 384-well microtiter plate was added 40 nL 200×test compound in DMSO and 4 μL 2× terbium chelate-labeled menin (videinfra for preparation) in assay buffer (40 mM Tris.HCl, pH 7.5, 50 mMNaCl, 1 mM DTT and 0.05% Pluronic F-127). After incubation of testcompound and terbium chelate-labeled menin for 5 min at ambienttemperature, 4 μL 2× FITC-MBM1 peptide (FITC-β-alanine-SARWRFPARPGT-NH₂)in assay buffer was added, the microtiter plate centrifuged at 1000 rpmfor 1 min and the assay mixtures incubated for 15 min at ambienttemperature. The relative amount of menin FITC-MBM1 complex present inan assay mixture is determined by measuring the homogenous time-resolvedfluorescence (HTRF) of the terbium/FITC donor/acceptor fluorphore pairusing a BMG Pherastar plate reader (ex. 337 nm/terbium em. 490 nm/FITCem. 520 nm) at ambient temperature. The degree of fluorescence resonanceenergy transfer (the HTRF value) is expressed as the ratio of thefluorescence emission intensities of the FITC and terbium fluorophores(F^(em) 520 nm/F^(em) 490 nm). The final concentrations of reagents inthe binding assay are 100 μM terbium chelate-labeled menin, 75 nMFITC-MBM1 peptide and 0.5% DMSO in assay buffer. Dose-responsetitrations of test compounds are conducted using an 11 point, three-foldserial dilution scheme, starting at 31 μM.

Compound potencies were determined by first calculating % inhibition ateach compound concentration according to equation 1:

% inhibition=((HC−LC)−(HTRF ^(compound) −LC))/(HC−LC))*100  (Eqn 1)

Where LC and HC are the HTRF values of the assay in the presence orabsence of a saturating concentration of a compound that competes withFITC-MBM1 for binding to menin, and HTRF^(compound) is the measured HTRFvalue in the presence of the test compound. HC and LC HTRF valuesrepresent an average of at least 16 replicates per plate. For each testcompound, % inhibition values were plotted vs. the logarithm of the testcompound concentration, and the IC₅₀ value derived from fitting thesedata to equation 2:

% inhibition=Bottom+(Top−Bottom)/(1+10{circumflex over ( )}((logIC₅₀−log[cmpd])*h))  (Eqn 2)

Where Bottom and Top are the lower and upper asymptotes of thedose-response curve, respectively, IC₅₀ is the concentration of compoundthat yields 50% inhibition of signal and h is the Hill coefficient.

Preparation of Terbium cryptate labeling of Menin: Menin (a.a.1-610-6×his tag) was labeled with terbium cryptate as follows. 2 mg ofMenin was buffer exchanged into 1× phosphate buffered saline. 16 uMMenin was incubated with 4-fold molar excess NHS-terbium cryptate(Cisbio Bioassays, Bedford, Mass.) for 2 hours at room temperature.

The labeled protein was purified away from free label by running thereaction over a Superdex 200 Increase 10/300 GL column at 0.75 ml/min.Peak fractions were collected, aliquoted and frozen at −80° C.

MENIN Protein Sequence (SEQ ID NO: 1):MGLKAAQKTLFPLRSIDDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVIPTNVPELTFQPSPAPDPPGGLTYFPVADLSIIAALYARFTAQIRGAVDLSLYPREGGVSSRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGTKLDSSGVAFAVVGACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTVNAGVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLWLLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIASAKTYYRDEHIYPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVANDVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKWEEGSPTPVLHVGWATFLVQSLGRFEGQVRQKVRIVSREAEAAEAEEPWGEEAREGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPEGGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQVQMKKQKVSTPSDYTL SFLKRQRKGLHHHHHH

TABLE 4 Biological data in the Menin fluorescence polarization (FP)assay (1), Menin/MUL homogenous time-resolved fluorescence (HTRF) assay(3) and proliferation assay (2). Co. No. means compound number. Thevalues in table 4 are averaged values over all measurements. (1) (3) (2)(2) (2) (2) Menin Menin Spheroid Spheroid Spheroid Spheroid FP HTRFassay assay assay assay assay assay MV-4-11 MOLM14 K562 KG1 Co. (IC₅₀(IC₅₀ (IC₅₀ (IC₅₀ (IC₅₀ (IC₅₀ No. (μM)) (μM)) (μM)) (μM)) (μM)) (μM)) 18 0.14 227 3.9 7.7 >15  1 0.056 144 3.7 4.6 >15  3 0.10 216 3.8 5.6~15  14 0.11 178 2.2 4.9  20a 0.076 13 0.96 5 >15  21a 0.49 745 6.912.1 >15  21b 0.13 389 4.2 4.6 >15  19 0.81 1960  5 0.67 600 6.430.6 >15  12 0.63 2247 >15 >15 >15  13 0.042 39 0.73 4.2 >15  11 0.762374  10 2.31  9 0.84 1579  8 0.79 2525  6 0.24 339 2.4 10.1 >15  7 0.731216  4 0.73 3164  2 0.39 706 4.9 14.6 >15  15 0.57 471 14.0 >15 >15  220.069 12 0.55 2.0 >15  23 0.95 1208  24 0.49 1486 9.1 >15  25 0.31 14653.8 7.2  26 0.16 143 10.4 >15 197 0.30 541 7.3 >15  86 0.092 102 2.611.3 >15 133 0.12 385 2.1 >15  30 0.53 839 9.2 >15  88 0.32 486 >15 >15 89 0.045 31 1.3 10.5  90 0.11 288 11.1 >15  74 0.12 211 2.0 6.1  752.36  34 0.72 902 198 0.14 178 5.1 12.4 199 0.41 1196 12.9 >15  35 0.992375  36 0.4 777  37 0.17 177 1.5 3.1  76 0.68 901  40 0.84 1465 1951.07 1536  41 0.63 950 273 0.021 8 0.08 0.86 10.3 12 200 0.11 190 3.410.3 201 1075 12.7 >15 202 490 >15 137 1097 10.1 >15  44 344 4.1 >15  829 0.38 1.8 >15 >15 105 15 0.67 3.4 >15 108 753 >15 109 571 8.1  45 3303.4  46 448 4.1  47 669  48 483 2.0  49 310 2.1  50 665 110 236 4.0 111394 9.3  77 191 12.1  54 342 3.7  56 304 4.9 112 1540 274 7 0.362.3 >15 >15 203 422 7.2 145 37 1.7 7.3 146 46 2.4 147 341 8.2  57 1794.2 8.6 148 5 0.33 211 13 0.68 3.5 12.5 213 149 1.9 214 17 0.76 1.4 >15150 60 2.0 220 75 0.65 4.7 >15 >15 221 1157 222 4 0.2 223 496 6.1 228238  58 185 2.7  59 1245 230 44 231 107 232 914 233 782 234 1045 235 230.71 236 1041 237 52 1.2 238 659 154 328 155 69 0.7 1.2 >15 243 249 244848 245 10 0.32 2 >15 246 284 247 19 0.45 2.1 >15 >15 248 549  61 513249 912 250 190 251 1660 252 1007 253 323 254 861 156 21 0.79 157 101158 281 159 1174  62 28 0.64 4 >15 161 187 4.2 258 3083 193 25 0.651.7 >15 >15 162 77 3.1  63 201 196 4 0.3 1.1 >15 >15 163 277 164 1125259 20 1.3  64 1139  65 65 3.2 166 79 2.4 167 1126 168 3 0.19 0.86 11.915 169 176 3.5 170 138 4.3  84 20 0.58 1.5 >15 171 159 3.1  66 26 0.950.69 >15 172 124 2.9 173 96 3.2 174 1386  67 108 4.9 179 95 1.3 180 1301.9  69 186 1.2 182 61 0.78 1.9 >15 183 103 1.3  71 48 1.1 184 730.97 >15 186 91 1.3 187 72 1.4 269 8 0.67 4.6 >15 188 115 1.5 189 1080.92 >15 270 499 271 623 190 524 191 1263 192 619 312 339 313 159 3.6315 21 0.32 1.5 >15 316 20 0.4 0.96 >3.7 288 68 1.4 289 6 0.21 1.3 29025 0.47 1.2 291 13 0.66 1.2 >15 292 12 0.29 1.5 >15 294 82 1.83 293 811.67 307 11 0.28 1.5 10.7 299 57 3.18 295 34 1.16 6.7 303 6 0.32 1.02296 200 4.89 309 16 0.74 1.6 297 242  304b 440  304a 15 0.44 1.3 >15 317248 0.77 >15 318 72 1.09 298 122 4.23 314 25 0.73 320 259 1.37 321 2101.1 322 200 1.7 323 47 0.75 324 143 1.9  31 0.087 116 1.3 9.8 224 2365185 161 1.3 311 85 2.4

TABLE 5 Biological data in the Menin fluorescence polarization (FP)assay (1), Menin/MLL homogenous time-resolved fluorescence (HTRF) assay(3) and proliferation assay (2). Co. No. means compound number. Thevalues in table 5 are values for individual measurements (not averaged):in case a value was determined more than 1 time, each value is reportedindividually in Table 5. (1) (3) (2) (2) (2) (2) Menin Menin SpheroidSpheroid Spheroid Spheroid FP HTRF assay assay assay assay assay assayMV-4-11 MOLM14 K562 KG1 Co. (IC₅₀ (IC₅₀ (IC₅₀ (IC₅₀ (IC₅₀ (IC₅₀ No.(μM)) (μM)) (μM)) (μM)) (μM)) (μM))  20b 0.038   7  0.51 >15    >15 >15 2.5  16 0.134  63 4.7 9.3 >15 >15    8.7  17 0.054  14 4.9 3.4 11  0.762.3  0.84  28 0.517 1262 0.5 3.2 10.8  >15    10.9   29 0.346 1438 1.810.8  >15    >15    9.9  87 0.048  28 4.1 8.0 >15  0.63 2.8  0.58 4.6 0.63  0.68  0.52 138  62 ~15.4  7   1.0 1.4

1. A compound of Formula (I)

or a tautomer or a stereoisomeric form thereof, wherein R¹ is selectedfrom the group consisting of CH₃, CH₂F, CHF₂, and CF₃; R² is selectedfrom the group consisting of hydrogen and CH₃; L¹ represents a 7- to10-membered saturated spiroheterobicyclic system containing one or twoN-atoms provided that it is N-linked to the thienopyrimidinylheterocycle; and --L²-R³ is selected from (a), (b), (c), (d), (e), (f)or (g), wherein (a) L² is selected from the group consisting of >SO₂,>CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—; wherein (i) when L² is linked to acarbon atom of L¹, then R^(4a) and R⁵ are each independently selectedfrom the group consisting of hydrogen; —OR⁶; —NR^(7a)R^(7b);—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen, oxygen or sulfur atom; (ii) when L² islinked to a nitrogen atom of L¹, then R^(4a) is selected from the groupconsisting of hydrogen; —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionallysubstituted with a substituent selected from the group consisting offluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and C-linked 4- to 7-memberednon-aromatic heterocyclyl containing at least one nitrogen, oxygen orsulfur atom; R⁵ is selected from the group consisting of hydrogen; —OR⁶;—NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of fluoro, —CN,—OR⁸, and —NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromaticheterocyclyl containing at least one nitrogen, oxygen or sulfur atom;R^(4b) is selected from the group consisting of hydrogen and methyl; orR^(4a) and R^(4b) together with the carbon atom to which they areattached form a C₃₋₅cycloalkyl or a C-linked 4- to 6-memberedheterocyclyl containing an oxygen atom; wherein R⁶, R^(7a), R^(7b), R⁸,R^(9a) and R^(9b) are each independently selected from the groupconsisting of hydrogen; C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, and—C(═O)NR^(10a)R^(10b); and C₂₋₄alkyl substituted with a substituentselected from the group consisting of —OR¹¹ and —NR^(10a)R^(10b);wherein R^(10a), R^(10b) and R¹¹ are each independently selected fromthe group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to7-membered non-aromatic heterocyclyl containing at least one nitrogen,oxygen or sulfur atom; and R³ is selected from the group consisting ofAr, Het¹, Het², and a 7- to 10-membered saturated spirocarbobicyclicsystem; or (c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, twoor three fluoro substituents; or (d) L² is O and R³ is selected from thegroup consisting of C₃₋₆alkyl optionally substituted with one, two orthree fluoro substituents; Ar; Het¹; Het²; a 7- to 10-membered saturatedspirocarbobicyclic system; —CH₂—Ar; —CH₂-Het¹; —CH₂-Het²; and —CH₂-(a 7-to 10-membered saturated spirocarbobicyclic system); when L² is linkedto a carbon atom of L¹; or (e) --L²-R³ is —O—CHR⁵—R³ when L² is linkedto a carbon atom of L¹, wherein R⁵ is selected from the group consistingof —C(═O)NR^(13a)R^(13b); C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —OR¹⁴, and—NR^(15a)R^(15b); and C-linked 4- to 7-membered non-aromaticheterocyclyl containing at least one nitrogen, oxygen or sulfur atom;wherein R^(13a), R^(13b), R¹⁴, R^(15a) and R^(15b) are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro and —C(═O)NR^(16a)R^(16b); and C₂₋₄alkylsubstituted with a substituent selected from the group consisting of—OR¹⁷ and —NR^(16a)R^(16b); wherein R^(16a), R^(16b) and R¹⁷ are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyl;and C-linked 4- to 7-membered non-aromatic heterocyclyl containing atleast one nitrogen, oxygen or sulfur atom; and R³ is selected from thegroup consisting of hydrogen; C₁₋₄alkyl optionally substituted with one,two, or three fluoro substituents; —CN; Ar, Het¹; Het²; and a 7- to10-membered saturated spirocarbobicyclic system; or (g) --L²-R³ is

 and wherein Ar is phenyl or naphthyl, each of which may be optionallysubstituted with one, two, or three substituents each independentlyselected from the group consisting of halo, —CN, —OR²⁴,—NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); Het¹ is a monocyclicheteroaryl selected from the group consisting of pyridyl, 4-, 5- or6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl,pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl, thiadiazolyl,and isoxazolyl; or a bicyclic heteroaryl selected from the groupconsisting of imidazothiazolyl, imidazoimidazolyl, benzofuranyl,benzothiophenyl, benzimidazolyl, benzoxazolyl, isobenzoxazolyl,benzisoxazolyl, benzothiazolyl, benzisothiazolyl, isobenzofuranyl,indolyl, isoindolyl, indolizinyl, indolinyl, isoindolinyl, indazolyl,pyrazolopyridinyl, pyrazolopyrimidinyl, imidazopyridinyl,imidazopyrazinyl, imidazopyridazinyl; each of which may be optionallysubstituted with one, two, or three substituents each independentlyselected from the group consisting of halo, —CN, —OR²⁴,—NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(21b); and Het² is a non-aromaticheterocyclyl optionally substituted with one, two, or three substituentseach independently selected from the group consisting of halo, —CN,—OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); wherein R²⁴, R^(25a),R^(25b), R²⁶, R^(27a), and R^(27b) are each independently selected fromthe group consisting of hydrogen; C₁₋₄alkyl optionally substituted witha substituent selected from the group consisting of fluoro and—C(═O)NR^(28a)R^(28b); and C₂₋₄alkyl substituted with a substituentselected from the group consisting of —OR²⁹ and —NR^(28a)R^(28b);wherein R^(28a), R^(28b) and R²⁹ are each independently selected fromthe group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to7-membered non-aromatic heterocyclyl containing at least one nitrogen,oxygen or sulfur atom; or a pharmaceutically acceptable salt or asolvate thereof; provided that the following compounds, andpharmaceutically acceptable addition salts, and solvates thereof areexcluded:


2. The compound according to claim 1, wherein (a) L² is selected fromthe group consisting of >SO₂, >CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—;wherein (i) when L² is linked to a carbon atom of L¹, then R^(4a) and R⁵are each independently selected from the group consisting of hydrogen;—OR⁶; —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionallysubstituted with a substituent selected from the group consisting offluoro, —CN, —OR⁸, and —N^(9a)R^(9b); and C-linked 4- to 7-memberednon-aromatic heterocyclyl containing at least one nitrogen, oxygen orsulfur atom; (ii) when L² is linked to a nitrogen atom of L¹, thenR^(4a) is selected from the group consisting of hydrogen;—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen, oxygen or sulfur atom; R⁵ is selectedfrom the group consisting of hydrogen; —OR⁶; —NR^(7a)R^(7b);—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen, oxygen or sulfur atom; R^(4b) isselected from the group consisting of hydrogen and methyl; or>CR^(4a)R^(4b) form a >C₃₋₅cycloalkanediyl or a >C-linked 4- to6-membered heterocyclediyl containing an oxygen atom; wherein R⁶,R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each independently selectedfrom the group consisting of hydrogen; C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of fluoro, —CN,and —C(═O)NR^(10a)R^(10b); and C₂₋₄alkyl substituted with a substituentselected from the group consisting of —OR¹¹ and —NR^(10a)R^(10b);wherein R^(10a), R^(10b) and R¹¹ are each independently selected fromthe group consisting of hydrogen; and C₁₋₄alkyl; and R³ is selected fromthe group consisting of Ar, Het¹, Het², and a 7- to 10-memberedsaturated spirocarbobicyclic system; or (c) --L²-R³ is C₁₋₆alkyloptionally substituted with one, two or three fluoro substituents; or(d) L² is O and R³ is selected from the group consisting of Ar, Het¹;—CH₂—Ar, —CH₂-Het¹, and —CH₂-(a 7- to 10-membered saturatedspirocarbobicyclic system); when L² is linked to a carbon atom of L¹; or(e) --L²-R³ is selected from the group consisting of

wherein R¹⁸ is hydrogen; (f) and wherein Ar is phenyl optionallysubstituted with one, two, or three substituents each independentlyselected from the group consisting of halo, —CN, and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b); Het¹ is a monocyclic heteroaryl selected from thegroup consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl,pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclicheteroaryl selected from imidazopyridinyl, in particularimidazo[1,2-a]pyridinyl; each of which may be optionally substitutedwith one, two, or three substituents each independently selected fromthe group consisting of halo, —CN, and C₁₋₄alkyl optionally substitutedwith a substituent selected from the group consisting of fluoro, —CN,—OR²⁶, —NR^(27a)R^(27b), and —C(═O)NR^(27a)R²⁷; and Het² is anon-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl andpiperidinyl; wherein R²⁶, R^(27a), and R^(27b) are each independentlyselected from the group consisting of hydrogen and C₁₋₄alkyl.
 3. Thecompound according to claim 1, wherein R¹ is CF₃; (a) L² is>CR^(4a)R^(4b); wherein R^(4a) is selected from the group consisting ofhydrogen; —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl; and C-linked 4- to 7-memberednon-aromatic heterocyclyl containing at least one nitrogen, oxygen orsulfur atom; and R^(4b) is selected from the group consisting ofhydrogen and methyl; wherein R^(7a) and R^(7b) are each independentlyselected from the group consisting of hydrogen; C₁₋₄alkyl; and C₂₋₄alkylsubstituted with a substituent selected from the group consisting of—OR¹¹ and —R^(10a)R^(10b); wherein R^(10a), R^(10b) and R¹¹ are eachindependently selected from the group consisting of hydrogen andC₁₋₄alkyl; and R³ is selected from the group consisting of Ar, Het¹,Het², and a 7- to 10-membered saturated spirocarbobicyclic system; or(c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two or threefluoro substituents; or (d) L² is O and R³ is selected from the groupconsisting of Ar, Het¹, —CH₂—Ar, —CH₂-Het¹, and —CH₂-(a 7- to10-membered saturated spirocarbobicyclic system); when L² is linked to acarbon atom of L¹; or (e) --L²-R³ is selected from the group consistingof

wherein R¹⁸ is hydrogen; and wherein Ar is phenyl optionally substitutedwith a halo substituent; Het¹ is a monocyclic heteroaryl selected fromthe group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl,pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, and 4- or 5-thiazolyl; ora bicyclic heteroaryl selected from imidazopyridinyl, in particularimidazo[1,2-a]pyridinyl; each of which may be optionally substitutedwith one, two, or three substituents each independently selected fromthe group consisting of halo and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(21b); and Het² is a non-aromaticheterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl;wherein R²⁶, R^(27a), and R^(27b) are each independently selected fromthe group consisting of hydrogen and C₁₋₄alkyl.
 4. The compoundaccording to claim 1, wherein R¹ is CF₃; L¹ represents a N-linked 7- to10-membered saturated spiroheterobicyclic system containing one or twoN-atoms selected from the group consisting of (a), (b), (c), (d), (e),(f) and (g)

wherein a represents the position of linkage to the thienopyrimidinylheterocycle; (a) L² is >CH₂; and R³ is selected from the groupconsisting of Ar, Het¹, and a 7- to 10-membered saturatedspirocarbobicyclic system; (b) and wherein Ar is phenyl optionallysubstituted with a halo substituent; and Het¹ is a monocyclic heteroarylselected from the group consisting of 4-, 5- or 6-pyrimidinyl,pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, and 4- or5-thiazolyl; or a bicyclic heteroaryl selected from imidazopyridinyl, inparticular imidazo[1,2-a]pyridinyl; each of which may be optionallysubstituted with one or two substituents each independently selectedfrom the group consisting of halo and C₁₋₄alkyl.
 5. The compoundaccording to claim 1, wherein R¹ is CF₃; R² is hydrogen; L¹ represents aN-linked 7- to 10-membered saturated spiroheterobicyclic systemcontaining one or two N-atoms selected from the group consisting of (a),(b), (c), (d), (e), (f) and (g)

wherein a represents the position of linkage to the thienopyrimidinylheterocycle; (a) L² is >CH₂; and R³ is selected from the groupconsisting of Ar, Het¹, and a 7- to 10-membered saturatedspirocarbobicyclic system; and wherein Ar is phenyl optionallysubstituted with a halo substituent; and Het¹ is a monocyclic heteroarylselected from the group consisting of 4-, 5- or 6-pyrimidinyl,pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, and imidazolyl; or abicyclic heteroaryl selected from imidazopyridinyl, in particularimidazo[1,2-a]pyridinyl; each of which may be optionally substitutedwith one or two substituents each independently selected from the groupconsisting of halo and C₁₋₄alkyl.
 6. The compound according to claim 1,wherein R¹ is CF₃; R² is selected from the group consisting of hydrogenand CH₃; L¹ represents a 7- to 10-membered saturated spiroheterobicyclicsystem containing one or two N-atoms provided that it is N-linked to thethienopyrimidinyl heterocycle; and --L²-R³ is selected from (a), (b),(c), (d), (f) or (g), wherein (a) L² is selected from the groupconsisting of >CR^(4a)R^(4b), and —CHR^(4a)CHR⁵—; wherein L² is linkedto a nitrogen atom of L¹; R^(4a) is selected from the group consistingof hydrogen; —C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted witha substituent selected from the group consisting of —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen atom; R⁵ is selected from the groupconsisting of hydrogen; —OR⁶; and C₁₋₄alkyl; R^(4b) is selected from thegroup consisting of hydrogen and methyl; or R^(4a) and R^(4b) togetherwith the carbon atom to which they are attached form a C₃₋₅cycloalkyl ora C-linked 4- to 6-membered heterocyclyl containing an oxygen atom;wherein R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are each independentlyselected from the group consisting of hydrogen; and C₂₋₄alkylsubstituted with a substituent selected from the group consisting of—OR¹¹ and —NR^(10a)R^(10b); wherein R^(10a), R^(10b) and R¹¹ are eachindependently selected from the group consisting of hydrogen; andC₁₋₄alkyl; and R³ is selected from the group consisting of Ar, Het¹,Het², and a 7- to 10-membered saturated spirocarbobicyclic system; (b)or (c) --L²-R³ is C₁₋₆alkyl optionally substituted with one, two orthree fluoro substituents; or (d) L² is O and R³ is —CH₂—Ar; or (g)--L²-R³ is

and wherein Ar is phenyl which may be optionally substituted with one,two, or three substituents each independently selected from the groupconsisting of halo, —OR²⁴, and C₁₋₄alkyl optionally substituted with—OR²⁶; Het¹ is a monocyclic heteroaryl selected from the groupconsisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl,pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl, andisoxazolyl; or a bicyclic heteroaryl selected from the group consistingof indolyl, imidazopyridinyl; each of which may be optionallysubstituted with one, two, or three substituents each independentlyselected from the group consisting of halo, —CN, —OR²⁴, and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of —CN, —OR²⁶, and —NR^(27a)R^(27b); and Het² is anon-aromatic heterocyclyl optionally substituted with one, two, or threesubstituents each independently selected from the group consisting ofhalo, —CN, and C₁₋₄alkyl optionally substituted with —OR²⁶; wherein R²⁴,R²⁶, R^(27a), and R^(27b) are each independently selected from the groupconsisting of hydrogen; C₁₋₄alkyl; and C₂₋₄alkyl substituted with—NR^(28a)R^(28b); wherein R^(28a) and R^(28b) are hydrogen.
 7. Thecompound according to claim 1, wherein R¹ is selected from the groupconsisting of CH₃, CH₂F, CHF₂, and CF₃; R² is selected from the groupconsisting of hydrogen and CH₃; L¹ represents a 7- to 10-memberedsaturated spiroheterobicyclic system containing one or two N-atomsprovided that it is N-linked to the thienopyrimidinyl heterocycle; and--L²-R³ is selected from (a), (b), (d), (e), or (f), wherein (a) L² isselected from the group consisting of >SO₂, >CR^(4a)R^(4b), and—CHR^(4a)CHR⁵—; wherein (i) when L² is linked to a carbon atom of L¹,then R^(4a) and R⁵ are each independently selected from the groupconsisting of hydrogen; —OR⁶; —NR^(7a)R^(7b); —C(═O)NR^(7a)R^(7b);C₁₋₄alkyl optionally substituted with a substituent selected from thegroup consisting of fluoro, —CN, —OR⁸, and —NR^(9a)R^(9b); and C-linked4- to 7-membered non-aromatic heterocyclyl containing at least onenitrogen, oxygen or sulfur atom; (ii) when L² is linked to a nitrogenatom of L¹, then R^(4a) is selected from the group consisting of—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen, oxygen or sulfur atom; R⁵ is selectedfrom the group consisting of hydrogen; —OR⁶; —NR^(7a)R^(7b);—C(═O)NR^(7a)R^(7b); C₁₋₄alkyl optionally substituted with a substituentselected from the group consisting of fluoro, —CN, —OR⁸, and—NR^(9a)R^(9b); and C-linked 4- to 7-membered non-aromatic heterocyclylcontaining at least one nitrogen, oxygen or sulfur atom; R^(4b) isselected from the group consisting of hydrogen and methyl; or R^(4a) andR^(4b) together with the carbon atom to which they are attached form aC₃₋₅cycloalkyl or a C-linked 4- to 6-membered heterocyclyl containing anoxygen atom; wherein R⁶, R^(7a), R^(7b), R⁸, R^(9a) and R^(9b) are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, and —C(═O)NR^(10a)R^(10b); and C₂₋₄alkylsubstituted with a substituent selected from the group consisting of—OR¹¹ and —NR^(10a)R^(10b); wherein R^(10a), R^(10b) and R¹¹ are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyl;and C-linked 4- to 7-membered non-aromatic heterocyclyl containing atleast one nitrogen, oxygen or sulfur atom; and R³ is selected from thegroup consisting of Ar, Het¹, Het², and a 7- to 10-membered saturatedspirocarbobicyclic system; or (d) L² is O and R³ is selected from thegroup consisting of C₃₋₆alkyl optionally substituted with one, two orthree fluoro substituents; Ar; Het¹; Het²; a 7- to 10-membered saturatedspirocarbobicyclic system; —CH₂—Ar; —CH₂-Het¹; —CH₂-Het²; and —CH₂-(a 7-to 10-membered saturated spirocarbobicyclic system); when L² is linkedto a carbon atom of L¹; or (e) --L²-R³ is —O—CHR⁵—R³ when L² is linkedto a carbon atom of L¹, wherein R⁵ is selected from the group consistingof —C(═O)NR^(13a)R^(13b); C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —OR¹⁴, and—NR^(15a)R^(15b); and C-linked 4- to 7-membered non-aromaticheterocyclyl containing at least one nitrogen, oxygen or sulfur atom;wherein R^(13a), R^(13b), R¹⁴, R^(15a) and R^(15b) are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro and —C(═O)NR^(16a)R^(16b); and C₂₋₄alkylsubstituted with a substituent selected from the group consisting of—OR¹⁷ and —NR^(16a)R^(16b); wherein R^(16a), R^(16b) and R¹⁷ are eachindependently selected from the group consisting of hydrogen; C₁₋₄alkyl;and C-linked 4- to 7-membered non-aromatic heterocyclyl containing atleast one nitrogen, oxygen or sulfur atom; and R³ is selected from thegroup consisting of hydrogen; C₁₋₄alkyl optionally substituted with one,two, or three fluoro substituents; —CN; Ar, Het¹; Het²; and a 7- to10-membered saturated spirocarbobicyclic system; and wherein Ar isphenyl or naphthyl, each of which may be optionally substituted withone, two, or three substituents each independently selected from thegroup consisting of halo of —CN, —OR²⁴, —NR^(25a)R^(25b), C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b); Het¹ is a monocyclic heteroaryl selected from thegroup consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl,pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclicheteroaryl selected from the group consisting of imidazothiazolyl,imidazoimidazolyl, benzofuranyl, benzothiophenyl, benzimidazolyl,benzoxazolyl, isobenzoxazolyl, benzisoxazolyl, benzothiazolyl,benzisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolizinyl,indolinyl, isoindolinyl, indazolyl, pyrazolopyridinyl,pyrazolopyrimidinyl, imidazopyridinyl, imidazopyrazinyl,imidazopyridazinyl; each of which may be optionally substituted withone, two, or three substituents each independently selected from thegroup consisting of halo, —CN, —OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyloptionally substituted with a substituent selected from the groupconsisting of fluoro, —CN, —OR²⁶, —NR^(27a)R^(27b), and—C(═O)NR^(27a)R^(27b); and Het² is a non-aromatic heterocyclyloptionally substituted with one, two, or three substituents eachindependently selected from the group consisting of halo, —CN, —OR²⁴,—NR^(25a)R^(23b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); wherein R²⁴, R^(25a),R^(25b), R²⁶, R^(27a), and R^(27b) are each independently selected fromthe group consisting of hydrogen; C₁₋₄alkyl optionally substituted witha substituent selected from the group consisting of fluoro and—C(═O)NR^(28a)R^(28b); and C₂₋₄alkyl substituted with a substituentselected from the group consisting of —OR²⁹ and —NR^(28a)R^(28b);wherein R^(28a), R^(28b) and R²⁹ are each independently selected fromthe group consisting of hydrogen; C₁₋₄alkyl; and C-linked 4- to7-membered non-aromatic heterocyclyl containing at least one nitrogen,oxygen or sulfur atom.
 8. The compound according to claim 1, wherein--L²-R³ is (a); R³ is Het¹ or Het²; Het¹ is a monocyclic heteroarylselected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl,pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, and imidazolyl; each ofwhich may be optionally substituted with one, two, or three substituentseach independently selected from the group consisting of halo, —CN,—OR²⁴, —NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b); and Het² is a non-aromaticheterocyclyl selected from the group consisting of azetidinyl,pyrrolidinyl, and piperidinyl; each of which may be optionallysubstituted with one, two, or three substituents each independentlyselected from the group consisting of halo, —CN, —OR²⁴,—NR^(25a)R^(25b), and C₁₋₄alkyl optionally substituted with asubstituent selected from the group consisting of fluoro, —CN, —OR²⁶,—NR^(27a)R^(27b), and —C(═O)NR^(27a)R^(27b).
 9. A pharmaceuticalcomposition comprising a compound as claimed in claim 1 and apharmaceutically acceptable carrier or diluent.
 10. A process forpreparing a pharmaceutical composition comprising mixing apharmaceutically acceptable carrier with a therapeutically effectiveamount of a compound according to claim
 1. 11. (canceled)
 12. (canceled)13. (canceled)
 14. (canceled)
 15. A method of treating a disorderselected from cancer, myelodysplastic syndrome (MDS) and diabetescomprising administering to a subject in need thereof, a therapeuticallyeffective amount of a compound as claimed in claim 1 or a pharmaceuticalcomposition comprising the compound.
 16. The method according to claim15 wherein the disorder is cancer.
 17. The method according to claim 16wherein cancer is selected from leukemias, myeloma or a solid tumorcancer such as prostate cancer, lung cancer, breast cancer, pancreaticcancer, colon cancer, liver cancer, melanoma and glioblastoma.
 18. Themethod according to claim 17 wherein the leukemia is selected from acuteleukemias, chronic leukemias, myeloid leukemias, myelogeneous leukemias,lymphoblastic leukemias, lymphocytic leukemias, Acute myelogeneousleukemias (AML), Chronic myelogenous leukemias (CML), Acutelymphoblastic leukemias (ALL), Chronic lymphocytic leukemias (CLL), Tcell prolymphocytic leukemias (T-PLL), Large granular lymphocyticleukemia, Hairy cell leukemia (HCL), MLL-rearranged leukemias, MLL-PTDleukemias, MLL amplified leukemias, MLL-positive leukemias, andleukemias exhibiting HOX/MEIS1 gene expression signatures.